doc_id	sent_index	relation_id	relation	trigger	trigger_offset	arg_num	arg_head	arg_protein	arg_domain	arg_site	arg_sugar	arg_head_offset	arg_base_np	arg_base_np_offset	arg_np	arg_np_offset	sent_text	PSource	NProtein	NID	SiteSource	SiteName
1400872	2	0	theme	polymerase	487:496	arg1	reaction					504:511	a polymerase chain reaction to obtain the eight exons encoding SHBG	485:551	a polymerase chain reaction to obtain the eight exons encoding SHBG	485:551	This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG.					
1400872	1	1	theme	48K	292:294	arg1	forms					269:273	three molecular weight forms	246:273	three molecular weight forms of 56K, 52K, and 48K	246:294	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	7	2	theme	CHO	1646:1648	arg1	cells					1650:1654	CHO cells	1646:1654	CHO cells	1646:1654	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	7	3	theme	sulfate-polyacrylamide	1423:1444	arg1	electrophoresis					1450:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	5	4	theme	Chinese	1084:1090	arg1	cells					1112:1116	Chinese hamster ovary (CHO) cells	1084:1116	Chinese hamster ovary (CHO) cells	1084:1116	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	4	5	from	position	889:896	arg1	site					856:859	an additional consensus site	832:859	an additional consensus site for N-glycosylation at this position	832:896	This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA.					
1400872	8	6	theme	additional	1674:1683	arg1	subunit					1685:1691	This additional subunit	1669:1691	This additional subunit	1669:1691	This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells.					
1400872	5	7	theme	expression	1044:1053	arg1	vector					1055:1060	the pRc/CMV expression vector	1032:1060	the pRc/CMV expression vector	1032:1060	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	7	8	theme	gel	1446:1448	arg1	electrophoresis					1450:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	0	9	theme	additional	84:93	arg1	chain					108:112	an additional carbohydrate chain	81:112	an additional carbohydrate chain	81:112	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	5	10	theme	ovary	1100:1104	arg1	cells					1112:1116	Chinese hamster ovary (CHO) cells	1084:1116	Chinese hamster ovary (CHO) cells	1084:1116	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	4	11	theme	consensus	846:854	arg1	site					856:859	an additional consensus site	832:859	an additional consensus site for N-glycosylation at this position	832:896	This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA.					
1400872	6	12	theme	those	1325:1329	arg1	serum					1286:1290	the serum	1282:1290	the serum of either a normal individual or those who produce an electrophoretic variant (98%)	1282:1374	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	7	13	theme	sodium	1408:1413	arg1	electrophoresis					1450:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	3	14	from	substitution	636:647	arg1	polypeptide					690:700	the SHBG polypeptide	681:700	the SHBG polypeptide	681:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	4	15	theme	additional	835:844	arg1	site					856:859	an additional consensus site	832:859	an additional consensus site for N-glycosylation at this position	832:896	This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA.					
1400872	9	16	theme	CHO	1950:1952	arg1	cells					1954:1958	CHO cells	1950:1958	CHO cells	1950:1958	Nevertheless, its steroid-binding affinity was equal to normal SHBG produced by CHO cells or SHBG in serum.					
1400872	4	17	theme	SHBG	960:963	arg1	DNA					979:981	a human SHBG complementary DNA	952:981	a human SHBG complementary DNA	952:981	This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA.					
1400872	8	18	theme	recombinant	1822:1832	arg1	SHBG					1834:1837	recombinant SHBG	1822:1837	recombinant SHBG	1822:1837	This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells.					
1400872	1	19	attach	isolated	131:138	arg1	individual					148:157	an individual	145:157	an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)	145:405	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	19	attach	isolated	131:138	arg1	variant					219:225	homozygous for a sex hormone-binding globulin (SHBG) variant	166:225	homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)	166:405	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	19	attach	isolated	131:138	arg2	DNA					123:125	Genomic DNA	115:125	Genomic DNA	115:125	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	7	20	theme	normal	1622:1627	arg1	SHBG					1629:1632	normal SHBG	1622:1632	normal SHBG produced by CHO cells or in serum	1622:1666	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	0	21	theme	carbohydrate	95:106	arg1	chain					108:112	an additional carbohydrate chain	81:112	an additional carbohydrate chain	81:112	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	3	22	theme	SHBG	685:688	arg1	polypeptide					690:700	the SHBG polypeptide	681:700	the SHBG polypeptide	681:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	3	23	from	residue	666:672	arg1	Asn					658:660	Asp --> Asn	650:660	Asp --> Asn	650:660	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	3	23	from	residue	666:672	arg1	substitution					636:647	an amino acid substitution	622:647	an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide	622:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	6	24	theme	individual	1311:1320	arg1	serum					1286:1290	the serum	1282:1290	the serum of either a normal individual or those who produce an electrophoretic variant (98%)	1282:1374	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	8	25	from	synthesis	1846:1854	arg1	cells					1863:1867	CHO cells	1859:1867	CHO cells	1859:1867	This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells.					
1400872	6	26	from	SHBG	1274:1277	arg1	serum					1286:1290	the serum	1282:1290	the serum of either a normal individual or those who produce an electrophoretic variant (98%)	1282:1374	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	5	27	theme	pRc/CMV	1036:1042	arg1	vector					1055:1060	the pRc/CMV expression vector	1032:1060	the pRc/CMV expression vector	1032:1060	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	7	28	theme	CHO	1517:1519	arg1	cells					1521:1525	CHO cells	1517:1525	CHO cells	1517:1525	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	1	29	theme	denaturing	325:334	arg1	conditions					336:345	denaturing conditions	325:345	denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)	325:405	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	29	theme	denaturing	325:334	arg1	electrophoresis					390:404	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	348:404	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	348:404	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	30	theme	homozygous	166:175	arg1	variant					219:225	homozygous for a sex hormone-binding globulin (SHBG) variant	166:225	homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)	166:405	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	30	theme	homozygous	166:175	arg1	individual					148:157	an individual	145:157	an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)	145:405	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	0	31	theme	Molecular	0:8	arg1	analyses					10:17	Molecular analyses	0:17	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.	0:113	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	1	32	theme	molecular	252:260	arg1	forms					269:273	three molecular weight forms	246:273	three molecular weight forms of 56K, 52K, and 48K	246:294	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	9	33	theme	normal	1926:1931	arg1	SHBG					1933:1936	normal SHBG	1926:1936	normal SHBG produced by CHO cells or SHBG in serum	1926:1975	Nevertheless, its steroid-binding affinity was equal to normal SHBG produced by CHO cells or SHBG in serum.					
1400872	4	34	theme	complementary	965:977	arg1	DNA					979:981	a human SHBG complementary DNA	952:981	a human SHBG complementary DNA	952:981	This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA.					
1400872	5	35	theme	complementary	996:1008	arg1	DNA					1010:1012	The mutated complementary DNA	984:1012	The mutated complementary DNA	984:1012	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	8	36	theme	complexity	1771:1780	arg1	degree					1761:1766	a greater degree	1751:1766	a greater degree of complexity	1751:1780	This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells.					
1400872	6	37	theme	normal	1304:1309	arg1	individual					1311:1320	a normal individual	1302:1320	a normal individual	1302:1320	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	1	38	theme	weight	262:267	arg1	forms					269:273	three molecular weight forms	246:273	three molecular weight forms of 56K, 52K, and 48K	246:294	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	7	39	theme	dodecyl	1415:1421	arg1	electrophoresis					1450:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	1408:1464	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	3	40	theme	Sequence	554:561	arg1	analysis					563:570	Sequence analysis	554:570	Sequence analysis of these exons	554:585	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	7	41	theme	60K	1542:1544	arg1	subunit					1546:1552	a 60K subunit	1540:1552	a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum	1540:1666	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	6	42	from	cells	1258:1262	arg1	serum					1286:1290	the serum	1282:1290	the serum of either a normal individual or those who produce an electrophoretic variant (98%)	1282:1374	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	6	43	theme	CHO	1254:1256	arg1	%					1267:1267	85%	1265:1267	85%	1265:1267	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	6	43	theme	CHO	1254:1256	arg1	cells					1258:1262	CHO cells	1254:1262	CHO cells (85%)	1254:1268	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	2	44	theme	oligonucleotide	458:472	arg1	primers					474:480	intron-specific oligonucleotide primers	442:480	intron-specific oligonucleotide primers	442:480	This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG.					
1400872	1	45	theme	sex	183:185	arg1	SHBG					213:216	SHBG	213:216	SHBG	213:216	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	45	theme	sex	183:185	arg1	globulin					203:210	a sex hormone-binding globulin	181:210	a sex hormone-binding globulin (SHBG)	181:217	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	0	46	theme	sex	30:32	arg1	globulin					50:57	a human sex hormone-binding globulin	22:57	a human sex hormone-binding globulin variant	22:65	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	3	47	theme	same	711:714	arg1	mutation					716:723	the same mutation	707:723	the same mutation	707:723	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	9	48	theme	steroid-binding	1888:1902	arg1	affinity					1904:1911	its steroid-binding affinity	1884:1911	its steroid-binding affinity	1884:1911	Nevertheless, its steroid-binding affinity was equal to normal SHBG produced by CHO cells or SHBG in serum.					
1400872	3	49	theme	Asp	650:652	arg1	Asn					658:660	Asp --> Asn	650:660	Asp --> Asn	650:660	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	3	49	theme	Asp	650:652	arg1	substitution					636:647	an amino acid substitution	622:647	an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide	622:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	1	50	theme	hormone-binding	187:201	arg1	SHBG					213:216	SHBG	213:216	SHBG	213:216	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	50	theme	hormone-binding	187:201	arg1	globulin					203:210	a sex hormone-binding globulin	181:210	a sex hormone-binding globulin (SHBG)	181:217	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	0	51	theme	human	24:28	arg1	globulin					50:57	a human sex hormone-binding globulin	22:57	a human sex hormone-binding globulin variant	22:65	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	1	52	theme	56K	278:280	arg1	forms					269:273	three molecular weight forms	246:273	three molecular weight forms of 56K, 52K, and 48K	246:294	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	8	53	theme	CHO	1859:1861	arg1	cells					1863:1867	CHO cells	1859:1867	CHO cells	1859:1867	This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells.					
1400872	1	54	theme	sulfate-polyacrylamide	363:384	arg1	conditions					336:345	denaturing conditions	325:345	denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)	325:405	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	54	theme	sulfate-polyacrylamide	363:384	arg1	electrophoresis					390:404	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	348:404	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	348:404	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	3	55	theme	point	598:602	arg1	mutation					604:611	a point mutation	596:611	a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide	596:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	8	56	theme	carbohydrate	1789:1800	arg1	structures					1802:1811	the carbohydrate structures	1785:1811	the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells	1785:1867	This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells.					
1400872	0	57	theme	globulin	50:57	arg1	variant					59:65	a human sex hormone-binding globulin variant	22:65	a human sex hormone-binding globulin variant	22:65	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	5	58	theme	CHO	1107:1109	arg1	cells					1112:1116	Chinese hamster ovary (CHO) cells	1084:1116	Chinese hamster ovary (CHO) cells	1084:1116	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	8	59	theme	greater	1753:1759	arg1	degree					1761:1766	a greater degree	1751:1766	a greater degree of complexity	1751:1780	This additional subunit is larger than the variant in serum and probably reflects a greater degree of complexity in the carbohydrate structures added to recombinant SHBG during synthesis in CHO cells.					
1400872	1	60	theme	gel	386:388	arg1	conditions					336:345	denaturing conditions	325:345	denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis)	325:405	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	1	60	theme	gel	386:388	arg1	electrophoresis					390:404	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	348:404	sodium dodecyl sulfate-polyacrylamide gel electrophoresis	348:404	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	3	61	theme	exons	581:585	arg1	analysis					563:570	Sequence analysis	554:570	Sequence analysis of these exons	554:585	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	5	62	theme	hamster	1092:1098	arg1	cells					1112:1116	Chinese hamster ovary (CHO) cells	1084:1116	Chinese hamster ovary (CHO) cells	1084:1116	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	0	63	theme	hormone-binding	34:48	arg1	globulin					50:57	a human sex hormone-binding globulin	22:57	a human sex hormone-binding globulin variant	22:65	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	1	64	theme	52K	283:285	arg1	forms					269:273	three molecular weight forms	246:273	three molecular weight forms of 56K, 52K, and 48K	246:294	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	6	65	theme	normal	1230:1235	arg1	SHBG					1237:1240	normal SHBG	1230:1240	normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%)	1230:1374	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	2	66	theme	intron-specific	442:456	arg1	primers					474:480	intron-specific oligonucleotide primers	442:480	intron-specific oligonucleotide primers	442:480	This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG.					
1400872	5	67	theme	mutated	988:994	arg1	DNA					1010:1012	The mutated complementary DNA	984:1012	The mutated complementary DNA	984:1012	The mutated complementary DNA was inserted into the pRc/CMV expression vector, and transfected into Chinese hamster ovary (CHO) cells.					
1400872	1	68	theme	Genomic	115:121	arg1	DNA					123:125	Genomic DNA	115:125	Genomic DNA	115:125	Genomic DNA was isolated from an individual who is homozygous for a sex hormone-binding globulin (SHBG) variant that resolves into three molecular weight forms of 56K, 52K, and 48K during electrophoresis under denaturing conditions (sodium dodecyl sulfate-polyacrylamide gel electrophoresis).					
1400872	7	69	theme	light	1585:1589	arg1	subunits					1597:1604	the heavy (52K) and light (48K) subunits	1565:1604	a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum	1540:1666	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	4	70	dep	introduced	933:942	arg1	confirm					906:912	confirm	906:912	to confirm its utilization	903:928	This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA.					
1400872	3	71	theme	acid	631:634	arg1	Asn					658:660	Asp --> Asn	650:660	Asp --> Asn	650:660	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	3	71	theme	acid	631:634	arg1	substitution					636:647	an amino acid substitution	622:647	an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide	622:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	7	72	theme	Western	1470:1476	arg1	blotting					1478:1485	Western blotting	1470:1485	Western blotting	1470:1485	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	4	73	theme	human	954:958	arg1	DNA					979:981	a human SHBG complementary DNA	952:981	a human SHBG complementary DNA	952:981	This mutation introduces an additional consensus site for N-glycosylation at this position, and to confirm its utilization we introduced it into a human SHBG complementary DNA.					
1400872	3	74	theme	-->	654:656	arg1	Asn					658:660	Asp --> Asn	650:660	Asp --> Asn	650:660	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	3	74	theme	-->	654:656	arg1	substitution					636:647	an amino acid substitution	622:647	an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide	622:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	3	75	theme	amino	625:629	arg1	Asn					658:660	Asp --> Asn	650:660	Asp --> Asn	650:660	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	3	75	theme	amino	625:629	arg1	substitution					636:647	an amino acid substitution	622:647	an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide	622:700	Sequence analysis of these exons revealed a point mutation encoding an amino acid substitution (Asp --> Asn) at residue 327 in the SHBG polypeptide, and the same mutation was identified in three siblings who also appear to be homozygous for this trait.					
1400872	2	76	theme	chain	498:502	arg1	reaction					504:511	a polymerase chain reaction to obtain the eight exons encoding SHBG	485:551	a polymerase chain reaction to obtain the eight exons encoding SHBG	485:551	This material was amplified using intron-specific oligonucleotide primers in a polymerase chain reaction to obtain the eight exons encoding SHBG.					
1400872	7	77	theme	heavy	1569:1573	arg1	subunits					1597:1604	the heavy (52K) and light (48K) subunits	1565:1604	a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum	1540:1666	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	0	78	dep	analyses	10:17	arg1	evidence					68:75	evidence	68:75	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.	0:113	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	7	79	theme	SHBG	1492:1495	arg1	variant					1497:1503	the SHBG variant	1488:1503	the SHBG variant produced by CHO cells	1488:1525	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	6	80	theme	electrophoretic	1346:1360	arg1	variant					1362:1368	an electrophoretic variant	1343:1368	an electrophoretic variant (98%)	1343:1374	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	6	80	theme	electrophoretic	1346:1360	arg1	%					1373:1373	98%	1371:1373	98%	1371:1373	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	0	81	theme	variant	59:65	arg1	analyses					10:17	Molecular analyses	0:17	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.	0:113	Molecular analyses of a human sex hormone-binding globulin variant: evidence for an additional carbohydrate chain.					
1400872	7	82	dep	heavy	1569:1573	arg1	48K					1592:1594	48K	1592:1594	48K	1592:1594	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	7	82	dep	heavy	1569:1573	arg1	52K					1576:1578	52K	1576:1578	52K	1576:1578	Furthermore, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, the SHBG variant produced by CHO cells consisted of a 60K subunit as well as the heavy (52K) and light (48K) subunits associated with normal SHBG produced by CHO cells or in serum.					
1400872	6	83	dep	less	1172:1175	arg1	%					1186:1186	54%	1184:1186	54%	1184:1186	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
1400872	6	84	theme	concanavalin	1198:1209	arg1	A					1211:1211	concanavalin A	1198:1211	concanavalin A	1198:1211	The product was secreted normally but proportionally less of it (54%) bound to concanavalin A when compared to normal SHBG produced by CHO cells (85%), or SHBG in the serum of either a normal individual or those who produce an electrophoretic variant (98%).					
19202066	0	0	theme	glucosamine	82:92	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.					
19202066	8	1	from	IKK	1523:1525	arg1	oncogenesis					1530:1540	oncogenesis	1530:1540	oncogenesis	1530:1540	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	2	2	theme	positive	322:329	arg1	loop					340:343	the positive feedback loop	318:343	the positive feedback loop between IKK-NF-kappaB and glycolysis	318:380	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	0	3	theme	beta-N-acetyl	68:80	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.					
19202066	3	4	theme	human	663:667	arg1	fibroblasts					669:679	transformed human fibroblasts	651:679	transformed human fibroblasts	651:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	7	5	theme	IKKbeta	1152:1158	arg1	O-GlcNAcylation					1133:1147	O-GlcNAcylation	1133:1147	O-GlcNAcylation of IKKbeta	1133:1158	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	5	6	theme	IKKbeta	902:908	arg1	O-GlcNAcylation					883:897	the O-GlcNAcylation	879:897	the O-GlcNAcylation of IKKbeta	879:908	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.					
19202066	8	7	theme	activity	1420:1427	arg1	enhancement					1395:1405	the enhancement	1391:1405	the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis	1391:1540	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	6	8	theme	O-GlcNAcase	979:989	arg1	streptozotocin					1001:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.					
19202066	8	9	theme	NF-kappaB	1410:1418	arg1	activity					1420:1427	NF-kappaB activity	1410:1427	NF-kappaB activity	1410:1427	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	7	10	theme	catalytic	1322:1330	arg1	activity					1332:1339	its catalytic activity	1318:1339	its catalytic activity	1318:1339	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	4	11	theme	activating	741:750	arg1	phosphorylation					752:766	the activating phosphorylation	737:766	the activating phosphorylation of IKK	737:773	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.					
19202066	8	12	theme	novel	1371:1375	arg1	mechanism					1377:1385	a novel mechanism	1369:1385	a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis	1369:1540	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	7	13	theme	phosphorylation	1246:1260	arg1	domain					1198:1203	the C-terminal domain	1183:1203	the C-terminal domain	1183:1203	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	7	13	theme	phosphorylation	1246:1260	arg1	site					1262:1265	an inactivating phosphorylation site	1230:1265	an inactivating phosphorylation site	1230:1265	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	4	14	theme	O-GlcNAcylated	710:723	arg1	IKKbeta					725:731	the O-GlcNAcylated IKKbeta	706:731	the O-GlcNAcylated IKKbeta	706:731	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.					
19202066	2	15	theme	IKK-NF-kappaB	263:275	arg1	pathway					277:283	the IKK-NF-kappaB pathway	259:283	the IKK-NF-kappaB pathway	259:283	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	1	16	theme	critical	159:166	arg1	role					168:171	a critical role	157:171	a critical role	157:171	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.					
19202066	3	17	theme	complex	502:508	arg1	IKKbeta					470:476	IKKbeta	470:476	IKKbeta	470:476	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	17	theme	complex	502:508	arg1	component					481:489	a component	479:489	a component of the IKK complex	479:508	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	8	18	theme	positive	1458:1465	arg1	regulation					1476:1485	positive feedback regulation	1458:1485	positive feedback regulation	1458:1485	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	5	19	theme	high	858:861	arg1	glucose					863:869	high glucose	858:869	high glucose	858:869	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.					
19202066	3	20	link	O-linked	544:551	arg1	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	20	link	O-linked	544:551	arg1	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	7	21	theme	IKKbeta	1284:1290	arg1	O-GlcNAcylation					1292:1306	IKKbeta O-GlcNAcylation	1284:1306	IKKbeta O-GlcNAcylation	1284:1306	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	5	22	theme	IKKbeta	947:953	arg1	activity					955:962	the TNFalpha-dependent IKKbeta activity	924:962	the TNFalpha-dependent IKKbeta activity	924:962	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.					
19202066	2	23	theme	cell	423:426	arg1	transformation					428:441	oncogene-induced cell transformation	406:441	oncogene-induced cell transformation	406:441	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	6	24	theme	inhibitor	991:999	arg1	streptozotocin					1001:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.					
19202066	3	25	theme	O-linked	544:551	arg1	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	25	theme	O-linked	544:551	arg1	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	7	26	theme	C-terminal	1187:1196	arg1	domain					1198:1203	the C-terminal domain	1183:1203	the C-terminal domain	1183:1203	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	7	26	theme	C-terminal	1187:1196	arg1	site					1262:1265	an inactivating phosphorylation site	1230:1265	an inactivating phosphorylation site	1230:1265	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	2	27	theme	oncogene-induced	406:421	arg1	transformation					428:441	oncogene-induced cell transformation	406:441	oncogene-induced cell transformation	406:441	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	4	28	theme	p65/NF-kappaB	793:805	arg1	knockdown					807:815	p65/NF-kappaB knockdown	793:815	p65/NF-kappaB knockdown	793:815	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.					
19202066	8	29	theme	feedback	1467:1474	arg1	regulation					1476:1485	positive feedback regulation	1458:1485	positive feedback regulation	1458:1485	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	3	30	from	glucosamine	567:577	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	30	from	glucosamine	567:577	arg1	fibroblasts					669:679	transformed human fibroblasts	651:679	transformed human fibroblasts	651:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	30	from	glucosamine	567:577	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	0	31	theme	p53	8:10	arg1	Loss					0:3	Loss	0:3	Loss of p53	0:10	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.					
19202066	3	32	theme	IKK	498:500	arg1	complex					502:508	the IKK complex	494:508	the IKK complex	494:508	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	1	33	theme	IkappaB	112:118	arg1	IKK					128:130	IKK	128:130	IKK	128:130	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.					
19202066	1	33	theme	IkappaB	112:118	arg1	kinase					120:125	IkappaB kinase	112:125	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.					
19202066	8	34	theme	p53	1440:1442	arg1	loss					1432:1435	loss	1432:1435	loss	1432:1435	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	2	35	theme	integral	389:396	arg1	role					398:401	an integral role	386:401	an integral role	386:401	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	4	36	theme	p53-deficient	685:697	arg1	cells					699:703	p53-deficient cells	685:703	p53-deficient cells	685:703	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.					
19202066	7	37	theme	inactivating	1233:1244	arg1	domain					1198:1203	the C-terminal domain	1183:1203	the C-terminal domain	1183:1203	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	7	37	theme	inactivating	1233:1244	arg1	site					1262:1265	an inactivating phosphorylation site	1230:1265	an inactivating phosphorylation site	1230:1265	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	3	38	mod	modified	530:537	arg1	IKKbeta					470:476	IKKbeta	470:476	IKKbeta	470:476	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	38	mod	modified	530:537	arg1	component					481:489	a component	479:489	a component of the IKK complex	479:508	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	38	mod	modified	530:537	arg3	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	38	mod	modified	530:537	arg3	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	0	39	theme	catalytic	21:29	arg1	activity					31:38	catalytic activity	21:38	catalytic activity of IKKbeta	21:49	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.					
19202066	3	40	theme	mouse	612:616	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	40	theme	mouse	612:616	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	6	41	theme	IKKbeta	1090:1096	arg1	O-GlcNAcylation					1028:1042	O-GlcNAcylation	1028:1042	O-GlcNAcylation	1028:1042	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.					
19202066	6	41	theme	IKKbeta	1090:1096	arg1	phosphorylation					1071:1085	concomitant activating phosphorylation	1048:1085	concomitant activating phosphorylation of IKKbeta	1048:1096	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.					
19202066	0	42	theme	IKKbeta	43:49	arg1	activity					31:38	catalytic activity	21:38	catalytic activity of IKKbeta	21:49	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.					
19202066	7	43	theme	Mutational	1099:1108	arg1	analysis					1110:1117	Mutational analysis	1099:1117	Mutational analysis	1099:1117	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.					
19202066	3	44	theme	embryonic	618:626	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	44	theme	embryonic	618:626	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	45	theme	beta-N-acetyl	553:565	arg1	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	45	theme	beta-N-acetyl	553:565	arg1	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	1	46	theme	kinase	120:125	arg1	pathway					143:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.					
19202066	2	47	contain	has	382:384	arg2	role					398:401	an integral role	386:401	an integral role	386:401	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	2	47	contain	has	382:384	arg1	loop					340:343	the positive feedback loop	318:343	the positive feedback loop between IKK-NF-kappaB and glycolysis	318:380	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	4	48	theme	glucose	820:826	arg1	depletion					828:836	glucose depletion	820:836	glucose depletion	820:836	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.					
19202066	0	49	link	O-linked	59:66	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.					
19202066	2	50	theme	p53	313:315	arg1	absence					302:308	the absence	298:308	the absence of p53	298:315	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	3	51	theme	transformed	651:661	arg1	fibroblasts					669:679	transformed human fibroblasts	651:679	transformed human fibroblasts	651:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	8	52	theme	enhanced	1492:1499	arg1	metabolism					1509:1518	enhanced glucose metabolism	1492:1518	enhanced glucose metabolism	1492:1518	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	4	53	theme	IKK	771:773	arg1	phosphorylation					752:766	the activating phosphorylation	737:766	the activating phosphorylation of IKK	737:773	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.					
19202066	4	53	theme	IKK	771:773	arg1	IKKbeta					725:731	the O-GlcNAcylated IKKbeta	706:731	the O-GlcNAcylated IKKbeta	706:731	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.					
19202066	3	54	theme	p53-deficient	598:610	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	3	54	theme	p53-deficient	598:610	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.					
19202066	5	55	theme	TNFalpha-dependent	928:945	arg1	activity					955:962	the TNFalpha-dependent IKKbeta activity	924:962	the TNFalpha-dependent IKKbeta activity	924:962	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.					
19202066	0	56	theme	O-linked	59:66	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.					
19202066	2	57	theme	glucose	232:238	arg1	metabolism					240:249	glucose metabolism	232:249	glucose metabolism	232:249	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	1	58	theme	-NF-kappaB	132:141	arg1	pathway					143:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.					
19202066	6	59	theme	activating	1060:1069	arg1	phosphorylation					1071:1085	concomitant activating phosphorylation	1048:1085	concomitant activating phosphorylation of IKKbeta	1048:1096	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.					
19202066	8	60	theme	glucose	1501:1507	arg1	metabolism					1509:1518	enhanced glucose metabolism	1492:1518	enhanced glucose metabolism	1492:1518	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.					
19202066	2	61	theme	feedback	331:338	arg1	loop					340:343	the positive feedback loop	318:343	the positive feedback loop between IKK-NF-kappaB and glycolysis	318:380	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.					
19202066	6	62	theme	concomitant	1048:1058	arg1	phosphorylation					1071:1085	concomitant activating phosphorylation	1048:1085	concomitant activating phosphorylation of IKKbeta	1048:1096	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.					
16474139	4	0	theme	O-linked	896:903	arg1	oligosaccharides					905:920	O-linked oligosaccharides	896:920	O-linked oligosaccharides	896:920	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	3	1	theme	coronavirus	480:490	arg1	proteins					494:501	the coronavirus M proteins	476:501	the coronavirus M proteins	476:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	9	2	theme	SARS-CoV	1538:1545	arg1	protein					1550:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.					
16474139	11	3	theme	serine	1836:1841	arg1	substitution					1820:1831	substitution	1820:1831	substitution of serine and threonine residues in the ectodomain of the 3a protein	1820:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	7	4	theme	different	1300:1308	arg1	ways					1310:1313	different ways	1300:1313	different ways	1300:1313	Pulse-chase analysis showed that both proteins were modified, although in different ways.					
16474139	11	5	theme	3a	1891:1892	arg1	protein					1894:1900	the 3a protein	1887:1900	the 3a protein	1887:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	1	6	theme	reading	177:183	arg1	frame					185:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame	111:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	10	7	dep	in	1719:1720	arg1	situ					1722:1725	situ	1722:1725	situ	1722:1725	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	3	8	theme	membrane	524:531	arg1	spanning					533:540	triple membrane spanning	517:540	triple membrane spanning	517:540	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	8	theme	membrane	524:531	arg1	they					508:511	they	508:511	they	508:511	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	1	9	theme	3a	191:192	arg1	protein					245:251	a structural protein	232:251	a structural protein	232:251	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	1	9	theme	3a	191:192	arg1	protein					194:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	3	10	theme	viral	655:659	arg1	proteins					672:679	viral structural proteins	655:679	viral structural proteins	655:679	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	5	11	theme	conserved	927:935	arg1	glycosylation					937:949	The conserved glycosylation	923:949	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them	923:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	4	12	theme	crucial	792:798	arg1	role					800:803	a crucial role	790:803	a crucial role	790:803	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	12	13	theme	O-glycosylated	1985:1998	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	12	13	theme	O-glycosylated	1985:1998	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	3	14	theme	same	551:554	arg1	topology					556:563	the same topology	547:563	the same topology	547:563	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	9	15	theme	sialic	1596:1601	arg1	acids					1603:1607	sialic acids	1596:1607	sialic acids	1596:1607	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.					
16474139	6	16	theme	T7	1172:1173	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.					
16474139	10	17	theme	3a	1678:1679	arg1	protein					1681:1687	the 3a protein	1674:1687	the 3a protein	1674:1687	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	1	18	theme	acute	122:126	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	1	18	theme	acute	122:126	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	4	19	theme	M	774:774	arg1	protein					776:782	The M protein	770:782	The M protein	770:782	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	12	20	theme	3a	1968:1969	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	12	20	theme	3a	1968:1969	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	6	21	theme	vaccinia	1157:1164	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.					
16474139	1	22	theme	syndrome	140:147	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	1	22	theme	syndrome	140:147	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	3	23	theme	S	730:730	arg1	proteins					732:739	the E and S proteins	720:739	proteins	732:739	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	0	24	theme	triple-spanning	67:81	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	24	theme	triple-spanning	67:81	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	24	theme	triple-spanning	67:81	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	3	25	theme	E	724:724	arg1	proteins					732:739	the E and S proteins	720:739	proteins	732:739	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	5	26	theme	coronavirus	958:968	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	3	27	contain	have	447:450	arg1	It					429:430	It	429:430	It	429:430	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	27	contain	have	447:450	arg2	similarities					460:471	several similarities	452:471	several similarities to the coronavirus M proteins	452:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	2	28	theme	known	367:371	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	6	29	theme	metabolic	1206:1214	arg1	labeling					1216:1223	metabolic labeling	1206:1223	metabolic labeling	1206:1223	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.					
16474139	5	30	theme	proteins	972:979	arg1	glycosylation					937:949	The conserved glycosylation	923:949	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them	923:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	12	31	gly	glycosylated	2100:2111	arg1	protein					2080:2086	the SARS-CoV M protein	2065:2086	the SARS-CoV M protein	2065:2086	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	12	32	theme	M	2078:2078	arg1	protein					2080:2086	the SARS-CoV M protein	2065:2086	the SARS-CoV M protein	2065:2086	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	3	33	dep	Golgi	632:636	arg1	predominantly					618:630	predominantly	618:630	predominantly	618:630	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	0	34	dep	proteins	92:99	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	34	dep	proteins	92:99	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	34	dep	proteins	92:99	arg1	Glycosylation					0:12	Glycosylation	0:12	Glycosylation of the severe acute respiratory syndrome	0:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	34	dep	proteins	92:99	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	2	35	theme	coronaviruses	414:426	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	8	36	theme	3a	1418:1419	arg1	protein					1421:1427	the 3a protein	1414:1427	the 3a protein	1414:1427	While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N-glycosidase enzyme.					
16474139	11	37	dep	serine	1836:1841	arg1	residues					1857:1864	residues	1857:1864	residues	1857:1864	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	12	38	dep	glycosylated	2100:2111	arg1	N					2098:2098	N	2098:2098	N	2098:2098	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	3	39	theme	similar	581:587	arg1	localizations					603:615	similar intracellular localizations	581:615	similar intracellular localizations	581:615	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	40	dep	spanning	533:540	arg1	i					505:505	i	505:505	i	505:505	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	40	dep	spanning	533:540	arg1	both					646:649	both	646:649	both	646:649	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	5	41	theme	3a	1008:1009	arg1	protein					1011:1017	the 3a protein	1004:1017	the 3a protein	1004:1017	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	0	42	theme	acute	28:32	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	9	43	contain	contain	1588:1594	arg2	acids					1603:1607	sialic acids	1596:1607	sialic acids	1596:1607	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.					
16474139	9	43	contain	contain	1588:1594	arg1	protein					1550:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.					
16474139	0	44	theme	syndrome	46:53	arg1	Glycosylation					0:12	Glycosylation	0:12	Glycosylation of the severe acute respiratory syndrome	0:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	5	45	gly	glycosylation	937:949	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	5	45	gly	glycosylation	937:949	arg1	resemblance					989:999	the resemblance	985:999	the resemblance of the 3a protein to them	985:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	10	46	theme	mutants	1783:1789	arg1	assay					1743:1747	an in situ O-glycosylation assay	1716:1747	an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants	1716:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	2	47	theme	genes	316:320	arg1	genes					316:320	the so-called group-specific genes	287:320	the so-called group-specific genes	287:320	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	2	47	theme	genes	316:320	arg1	one					280:282	one	280:282	one	280:282	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	10	48	theme	endoplasmic	1752:1762	arg1	mutants					1783:1789	endoplasmic reticulum-retained mutants	1752:1789	endoplasmic reticulum-retained mutants	1752:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	12	49	theme	M	2043:2043	arg1	proteins					2045:2052	the group 2 coronavirus M proteins	2019:2052	the group 2 coronavirus M proteins	2019:2052	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	3	50	theme	M	492:492	arg1	proteins					494:501	the coronavirus M proteins	476:501	the coronavirus M proteins	476:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	51	dep	have	576:579	arg1	Golgi					632:636	Golgi	632:636	Golgi	632:636	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	51	dep	have	576:579	arg1	iii					641:643	iii	641:643	iii	641:643	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	51	dep	have	576:579	arg1	ii					567:568	ii	567:568	ii	567:568	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	2	52	theme	sequence	333:340	arg1	homology					342:349	no sequence homology	330:349	no sequence homology with any of the known structural or group-specific proteins of coronaviruses	330:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	1	53	theme	structural	234:243	arg1	protein					245:251	a structural protein	232:251	a structural protein	232:251	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	1	53	theme	structural	234:243	arg1	protein					194:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	11	54	theme	protein	1894:1900	arg1	protein					1894:1900	the 3a protein	1887:1900	the 3a protein	1887:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	11	54	theme	protein	1894:1900	arg1	ectodomain					1873:1882	the ectodomain	1869:1882	the ectodomain of the 3a protein	1869:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	2	55	theme	so-called	291:299	arg1	genes					316:320	the so-called group-specific genes	287:320	the so-called group-specific genes	287:320	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	9	56	theme	3a	1547:1548	arg1	protein					1550:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.					
16474139	8	57	theme	N-glycosidase	1512:1524	arg1	enzyme					1526:1531	the N-glycosidase enzyme	1508:1531	the N-glycosidase enzyme	1508:1531	While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N-glycosidase enzyme.					
16474139	1	58	theme	open	172:175	arg1	frame					185:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame	111:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	3	59	theme	triple	517:522	arg1	spanning					533:540	triple membrane spanning	517:540	triple membrane spanning	517:540	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	59	theme	triple	517:522	arg1	they					508:511	they	508:511	they	508:511	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	1	60	theme	frame	185:189	arg1	protein					245:251	a structural protein	232:251	a structural protein	232:251	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	1	60	theme	frame	185:189	arg1	protein					194:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	4	61	link	O-linked	896:903	arg1	oligosaccharides					905:920	O-linked oligosaccharides	896:920	O-linked oligosaccharides	896:920	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	3	62	theme	structural	661:670	arg1	proteins					672:679	viral structural proteins	655:679	viral structural proteins	655:679	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	63	dep	appear	696:701	arg1	iv					687:688	iv	687:688	iv	687:688	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	11	64	theme	threonine	1847:1855	arg1	substitution					1820:1831	substitution	1820:1831	substitution of serine and threonine residues in the ectodomain of the 3a protein	1820:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	11	65	theme	sugars	1941:1946	arg1	addition					1916:1923	the addition	1912:1923	the addition of the O-linked sugars	1912:1946	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	8	66	theme	M	1326:1326	arg1	protein					1328:1334	the M protein	1322:1334	the M protein	1322:1334	While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N-glycosidase enzyme.					
16474139	3	67	contain	have	576:579	arg2	localizations					603:615	similar intracellular localizations	581:615	similar intracellular localizations	581:615	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	3	67	contain	have	576:579	arg1	they					571:574	they	571:574	they	571:574	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	6	68	theme	expression	1175:1184	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.					
16474139	10	69	theme	protein	1681:1687	arg1	O-glycosylation					1655:1669	O-glycosylation	1655:1669	O-glycosylation of the 3a protein	1655:1687	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	1	70	theme	severe	115:120	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	1	70	theme	severe	115:120	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	11	71	from	substitution	1820:1831	arg1	protein					1894:1900	the 3a protein	1887:1900	the 3a protein	1887:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	11	71	from	substitution	1820:1831	arg1	ectodomain					1873:1882	the ectodomain	1869:1882	the ectodomain of the 3a protein	1869:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	6	72	theme	virus	1166:1170	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.					
16474139	10	73	theme	in	1719:1720	arg1	assay					1743:1747	an in situ O-glycosylation assay	1716:1747	an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants	1716:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	1	74	theme	respiratory	128:138	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	1	74	theme	respiratory	128:138	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	7	75	theme	Pulse-chase	1226:1236	arg1	analysis					1238:1245	Pulse-chase analysis	1226:1245	Pulse-chase analysis	1226:1245	Pulse-chase analysis showed that both proteins were modified, although in different ways.					
16474139	12	76	theme	SARS-CoV	1959:1966	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	12	76	theme	SARS-CoV	1959:1966	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	0	77	theme	membrane	83:90	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	77	theme	membrane	83:90	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	77	theme	membrane	83:90	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	12	78	gly	O-glycosylated	1985:1998	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	12	78	gly	O-glycosylated	1985:1998	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	1	79	theme	coronavirus	149:159	arg1	frame					185:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame	111:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.					
16474139	5	80	theme	resemblance	989:999	arg1	glycosylation					937:949	The conserved glycosylation	923:949	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them	923:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	9	81	theme	oligosaccharides	1637:1652	arg1	presence					1625:1632	the presence	1621:1632	the presence of oligosaccharides	1621:1652	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.					
16474139	10	82	theme	reticulum-retained	1764:1781	arg1	mutants					1783:1789	endoplasmic reticulum-retained mutants	1752:1789	endoplasmic reticulum-retained mutants	1752:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	12	83	gly	glycoprotein	2000:2011	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	12	83	gly	glycoprotein	2000:2011	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	2	84	contain	has	326:328	arg1	protein					258:264	The protein	254:264	The protein	254:264	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	2	84	contain	has	326:328	arg2	homology					342:349	no sequence homology	330:349	no sequence homology with any of the known structural or group-specific proteins of coronaviruses	330:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	3	85	theme	several	452:458	arg1	similarities					460:471	several similarities	452:471	several similarities to the coronavirus M proteins	452:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	5	86	theme	M	970:970	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	4	87	gly	glycosylated	836:847	arg1	coronaviruses					856:868	all coronaviruses	852:868	all coronaviruses	852:868	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	4	87	gly	glycosylated	836:847	arg1	protein					776:782	The M protein	770:782	The M protein	770:782	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	4	87	gly	glycosylated	836:847	arg2	protein					776:782	The M protein	770:782	The M protein	770:782	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	11	88	link	O-linked	1932:1939	arg1	sugars					1941:1946	the O-linked sugars	1928:1946	the O-linked sugars	1928:1946	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	4	89	theme	coronavirus	808:818	arg1	assembly					820:827	coronavirus assembly	808:827	coronavirus assembly	808:827	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.					
16474139	5	90	gly	glycosylation	1053:1065	arg1	proteins					1098:1105	these two SARS-CoV membrane proteins	1070:1105	these two SARS-CoV membrane proteins	1070:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	11	91	theme	O-linked	1932:1939	arg1	sugars					1941:1946	the O-linked sugars	1928:1946	the O-linked sugars	1928:1946	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.					
16474139	12	92	theme	SARS-CoV	2069:2076	arg1	protein					2080:2086	the SARS-CoV M protein	2065:2086	the SARS-CoV M protein	2065:2086	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	0	93	theme	severe	21:26	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	2	94	theme	group-specific	387:400	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	0	95	theme	respiratory	34:44	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	10	96	gly	O-glycosylation	1655:1669	arg1	protein					1681:1687	the 3a protein	1674:1687	the 3a protein	1674:1687	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	0	97	gly	Glycosylation	0:12	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	10	98	theme	O-glycosylation	1727:1741	arg1	assay					1743:1747	an in situ O-glycosylation assay	1716:1747	an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants	1716:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.					
16474139	5	99	theme	membrane	1089:1096	arg1	proteins					1098:1105	these two SARS-CoV membrane proteins	1070:1105	these two SARS-CoV membrane proteins	1070:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	2	100	theme	structural	373:382	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	0	101	theme	coronavirus	55:65	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	101	theme	coronavirus	55:65	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	0	101	theme	coronavirus	55:65	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.					
16474139	5	102	theme	protein	1011:1017	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	5	102	theme	protein	1011:1017	arg1	resemblance					989:999	the resemblance	985:999	the resemblance of the 3a protein to them	985:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	3	103	theme	intracellular	589:601	arg1	localizations					603:615	similar intracellular localizations	581:615	similar intracellular localizations	581:615	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.					
16474139	12	104	theme	group	2023:2027	arg1	proteins					2045:2052	the group 2 coronavirus M proteins	2019:2052	the group 2 coronavirus M proteins	2019:2052	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	2	105	theme	group-specific	301:314	arg1	genes					316:320	the so-called group-specific genes	287:320	the so-called group-specific genes	287:320	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	5	106	theme	SARS-CoV	1080:1087	arg1	proteins					1098:1105	these two SARS-CoV membrane proteins	1070:1105	these two SARS-CoV membrane proteins	1070:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
16474139	2	107	with	homology	342:349	arg1	any					356:358	any	356:358	any	356:358	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.					
16474139	12	108	theme	coronavirus	2031:2041	arg1	proteins					2045:2052	the group 2 coronavirus M proteins	2019:2052	the group 2 coronavirus M proteins	2019:2052	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.					
16474139	5	109	theme	proteins	1098:1105	arg1	glycosylation					1053:1065	the glycosylation	1049:1065	the glycosylation of these two SARS-CoV membrane proteins	1049:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.					
25903135	8	0	theme	dimer-of-dimers	1154:1168	arg1	architecture					1170:1181	The dimer-of-dimers architecture	1150:1181	The dimer-of-dimers architecture	1150:1181	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	2	1	theme	signaling	364:372	arg1	processes					374:382	cell signaling processes	359:382	cell signaling processes	359:382	It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes.					
25903135	1	2	theme	system	257:262	arg1	development					264:274	nervous system development	249:274	nervous system development	249:274	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	5	3	theme	C-terminal	798:807	arg1	dimer					821:825	a C-terminal β-propeller dimer	796:825	a C-terminal β-propeller dimer at the tips	796:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	5	4	theme	β-propeller	809:819	arg1	dimer					821:825	a C-terminal β-propeller dimer	796:825	a C-terminal β-propeller dimer at the tips	796:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	1	5	dep	known	97:101	arg1	also					92:95	also	92:95	also	92:95	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	2	6	theme	receptors	339:347	arg1	number					288:293	a number	286:293	a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes	286:382	It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes.					
25903135	5	7	theme	parallel	745:752	arg1	coil					786:789	a parallel dimeric disulfide-linked coiled coil	743:789	a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips	743:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	1	8	gly	glycoprotein	222:233	arg1	glycoprotein					222:233	a highly expressed neuronal glycoprotein	194:233	a highly expressed neuronal glycoprotein important for nervous system development	194:274	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	5	9	link	disulfide-linked	762:777	arg1	coil					786:789	a parallel dimeric disulfide-linked coiled coil	743:789	a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips	743:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	2	10	theme	secreted	298:305	arg1	proteins					307:314	secreted proteins	298:314	secreted proteins	298:314	It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes.					
25903135	5	11	theme	dimeric	754:760	arg1	coil					786:789	a parallel dimeric disulfide-linked coiled coil	743:789	a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips	743:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	6	12	theme	combination	931:941	arg1	structure					869:877	our crystal structure	857:877	our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation	857:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	6	13	dep	exposed	1058:1064	arg1	outward					1050:1056	outward	1050:1056	outward	1050:1056	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	3	14	theme	distinctive	591:601	arg1	architecture					612:623	a distinctive V-shaped architecture	589:623	a distinctive V-shaped architecture	589:623	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	4	15	theme	dimeric	680:686	arg1	domains					699:705	two disulfide-linked dimeric N-terminal domains	659:705	two disulfide-linked dimeric N-terminal domains	659:705	The base of the "V" is formed by two disulfide-linked dimeric N-terminal domains.					
25903135	5	16	theme	disulfide-linked	762:777	arg1	coil					786:789	a parallel dimeric disulfide-linked coiled coil	743:789	a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips	743:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	6	17	theme	β-propeller	919:929	arg1	combination					931:941	β-propeller combination	919:941	β-propeller combination	919:941	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	3	18	dep	determined	517:526	arg1	Using					385:389	Using	385:389	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy	385:512	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	3	19	theme	solution	437:444	arg1	scattering					446:455	solution scattering	437:455	solution scattering	437:455	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	1	20	theme	olfactomedin	147:158	arg1	superfamily					178:188	the olfactomedin domain-containing superfamily	143:188	the olfactomedin domain-containing superfamily	143:188	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	5	21	theme	coiled	779:784	arg1	coil					786:789	a parallel dimeric disulfide-linked coiled coil	743:789	a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips	743:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	5	22	from	tips	834:837	arg1	dimer					821:825	a C-terminal β-propeller dimer	796:825	a C-terminal β-propeller dimer at the tips	796:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	3	23	theme	crystallography	420:434	arg1	approach					402:409	a combined approach	391:409	a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy	391:512	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	7	24	theme	member	1101:1106	arg1	myocilin					1108:1115	its family member myocilin	1090:1115	its family member myocilin	1090:1115	Similar to its family member myocilin, Olfm1 is stabilized by calcium.					
25903135	4	25	theme	V	643:643	arg1	"					644:644	the "V"	638:644	the "V"	638:644	The base of the "V" is formed by two disulfide-linked dimeric N-terminal domains.					
25903135	8	26	theme	other	1302:1306	arg1	members					1335:1341	several other olfactomedin domain family members	1294:1341	several other olfactomedin domain family members	1294:1341	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	1	27	theme	domain-containing	160:176	arg1	superfamily					178:188	the olfactomedin domain-containing superfamily	143:188	the olfactomedin domain-containing superfamily	143:188	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	3	28	theme	combined	393:400	arg1	approach					402:409	a combined approach	391:409	a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy	391:512	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	6	29	theme	segment	907:913	arg1	structure					869:877	our crystal structure	857:877	our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation	857:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	4	30	theme	N-terminal	688:697	arg1	domains					699:705	two disulfide-linked dimeric N-terminal domains	659:705	two disulfide-linked dimeric N-terminal domains	659:705	The base of the "V" is formed by two disulfide-linked dimeric N-terminal domains.					
25903135	2	31	theme	cell	320:323	arg1	receptors					339:347	cell surface-bound receptors	320:347	cell surface-bound receptors	320:347	It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes.					
25903135	1	32	theme	superfamily	178:188	arg1	member					133:138	a member	131:138	a member of the olfactomedin domain-containing superfamily	131:188	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	1	32	theme	superfamily	178:188	arg1	glycoprotein					222:233	a highly expressed neuronal glycoprotein	194:233	a highly expressed neuronal glycoprotein important for nervous system development	194:274	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	1	32	theme	superfamily	178:188	arg1	Olfactomedin-1					69:82	Olfactomedin-1	69:82	Olfactomedin-1 (Olfm1; also known as noelin and pancortin)	69:126	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	6	33	theme	top	1034:1036	arg1	faces					1038:1042	the β-propeller top faces	1018:1042	the β-propeller top faces	1018:1042	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	3	34	theme	scattering	446:455	arg1	approach					402:409	a combined approach	391:409	a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy	391:512	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	0	35	theme	V-shaped	21:28	arg1	Structure					58:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	Olfactomedin-1 Has a V-shaped Disulfide-linked Tetrameric Structure.					
25903135	6	36	theme	dimeric	993:999	arg1	arrangement					1001:1011	a disulfide-linked dimeric arrangement	974:1011	a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation	974:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	7	37	theme	family	1094:1099	arg1	myocilin					1108:1115	its family member myocilin	1090:1115	its family member myocilin	1090:1115	Similar to its family member myocilin, Olfm1 is stabilized by calcium.					
25903135	0	38	link	Disulfide-linked	30:45	arg1	Structure					58:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	Olfactomedin-1 Has a V-shaped Disulfide-linked Tetrameric Structure.					
25903135	2	39	theme	proteins	307:314	arg1	number					288:293	a number	286:293	a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes	286:382	It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes.					
25903135	8	40	theme	several	1294:1300	arg1	members					1335:1341	several other olfactomedin domain family members	1294:1341	several other olfactomedin domain family members	1294:1341	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	6	41	theme	disulfide-linked	976:991	arg1	arrangement					1001:1011	a disulfide-linked dimeric arrangement	974:1011	a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation	974:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	6	42	with	arrangement	1001:1011	arg1	faces					1038:1042	the β-propeller top faces	1018:1042	the β-propeller top faces	1018:1042	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	6	43	theme	coiled-coil	895:905	arg1	segment					907:913	a C-terminal coiled-coil segment	882:913	a C-terminal coiled-coil segment	882:913	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	2	44	theme	cell	359:362	arg1	processes					374:382	cell signaling processes	359:382	cell signaling processes	359:382	It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes.					
25903135	3	45	theme	electron	494:501	arg1	microscopy					503:512	electron microscopy	494:512	electron microscopy	494:512	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	0	46	theme	Tetrameric	47:56	arg1	Structure					58:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	Olfactomedin-1 Has a V-shaped Disulfide-linked Tetrameric Structure.					
25903135	6	47	from	arrangement	1001:1011	arg1	orientation					1066:1076	an outward exposed orientation	1047:1076	an outward exposed orientation	1047:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	3	48	theme	ultracentrifugation	469:487	arg1	approach					402:409	a combined approach	391:409	a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy	391:512	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	6	49	link	disulfide-linked	976:991	arg1	arrangement					1001:1011	a disulfide-linked dimeric arrangement	974:1011	a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation	974:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	6	50	dep	segment	907:913	arg1	Olfm1					944:948	Olfm1	944:948	Olfm1(coil-Olf)	944:958	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	6	50	dep	segment	907:913	arg1	coil-Olf					950:957	coil-Olf	950:957	coil-Olf	950:957	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	1	51	theme	expressed	203:211	arg1	glycoprotein					222:233	a highly expressed neuronal glycoprotein	194:233	a highly expressed neuronal glycoprotein important for nervous system development	194:274	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	0	52	theme	Disulfide-linked	30:45	arg1	Structure					58:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	Olfactomedin-1 Has a V-shaped Disulfide-linked Tetrameric Structure.					
25903135	3	53	link	disulfide-linked	557:572	arg1	tetramers					574:582	disulfide-linked tetramers	557:582	disulfide-linked tetramers	557:582	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	6	54	theme	β-propeller	1022:1032	arg1	faces					1038:1042	the β-propeller top faces	1018:1042	the β-propeller top faces	1018:1042	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	8	55	theme	members	1335:1341	arg1	conformation					1278:1289	the conformation	1274:1289	the conformation of several other olfactomedin domain family members	1274:1341	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	1	56	theme	neuronal	213:220	arg1	glycoprotein					222:233	a highly expressed neuronal glycoprotein	194:233	a highly expressed neuronal glycoprotein important for nervous system development	194:274	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	3	57	theme	full-length	533:543	arg1	Olfm1					545:549	full-length Olfm1	533:549	full-length Olfm1	533:549	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	6	58	theme	exposed	1058:1064	arg1	orientation					1066:1076	an outward exposed orientation	1047:1076	an outward exposed orientation	1047:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	8	59	theme	family	1328:1333	arg1	members					1335:1341	several other olfactomedin domain family members	1294:1341	several other olfactomedin domain family members	1294:1341	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	3	60	theme	x-ray	414:418	arg1	crystallography					420:434	x-ray crystallography	414:434	x-ray crystallography	414:434	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	3	61	theme	microscopy	503:512	arg1	approach					402:409	a combined approach	391:409	a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy	391:512	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	8	62	theme	domain	1321:1326	arg1	members					1335:1341	several other olfactomedin domain family members	1294:1341	several other olfactomedin domain family members	1294:1341	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	0	63	contain	Has	15:17	arg1	Olfactomedin-1					0:13	Olfactomedin-1	0:13	Olfactomedin-1	0:13	Olfactomedin-1 Has a V-shaped Disulfide-linked Tetrameric Structure.					
25903135	0	63	contain	Has	15:17	arg2	Structure					58:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	a V-shaped Disulfide-linked Tetrameric Structure	19:66	Olfactomedin-1 Has a V-shaped Disulfide-linked Tetrameric Structure.					
25903135	8	64	theme	olfactomedin	1308:1319	arg1	members					1335:1341	several other olfactomedin domain family members	1294:1341	several other olfactomedin domain family members	1294:1341	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	6	65	theme	crystal	861:867	arg1	structure					869:877	our crystal structure	857:877	our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation	857:1076	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	8	66	theme	clustering	1212:1221	arg1	receptors					1223:1231	clustering receptors	1212:1231	clustering receptors	1212:1231	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	1	67	dep	Olfactomedin-1	69:82	arg1	known					97:101	known	97:101	known as noelin and pancortin	97:125	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	3	68	theme	V-shaped	603:610	arg1	architecture					612:623	a distinctive V-shaped architecture	589:623	a distinctive V-shaped architecture	589:623	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	4	69	theme	"	644:644	arg1	base					630:633	The base	626:633	The base of the "V"	626:644	The base of the "V" is formed by two disulfide-linked dimeric N-terminal domains.					
25903135	5	70	theme	V	724:724	arg1	legs					726:729	the two V legs	716:729	the two V legs	716:729	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	8	71	from	role	1194:1197	arg1	receptors					1223:1231	clustering receptors	1212:1231	clustering receptors	1212:1231	The dimer-of-dimers architecture suggests a role for Olfm1 in clustering receptors to regulate signaling and sheds light on the conformation of several other olfactomedin domain family members.					
25903135	4	72	link	disulfide-linked	663:678	arg1	domains					699:705	two disulfide-linked dimeric N-terminal domains	659:705	two disulfide-linked dimeric N-terminal domains	659:705	The base of the "V" is formed by two disulfide-linked dimeric N-terminal domains.					
25903135	2	73	theme	surface-bound	325:337	arg1	receptors					339:347	cell surface-bound receptors	320:347	cell surface-bound receptors	320:347	It binds a number of secreted proteins and cell surface-bound receptors to induce cell signaling processes.					
25903135	1	74	theme	important	235:243	arg1	glycoprotein					222:233	a highly expressed neuronal glycoprotein	194:233	a highly expressed neuronal glycoprotein important for nervous system development	194:274	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.					
25903135	6	75	theme	C-terminal	884:893	arg1	segment					907:913	a C-terminal coiled-coil segment	882:913	a C-terminal coiled-coil segment	882:913	This agrees with our crystal structure of a C-terminal coiled-coil segment and β-propeller combination (Olfm1(coil-Olf)) that reveals a disulfide-linked dimeric arrangement with the β-propeller top faces in an outward exposed orientation.					
25903135	4	76	theme	disulfide-linked	663:678	arg1	domains					699:705	two disulfide-linked dimeric N-terminal domains	659:705	two disulfide-linked dimeric N-terminal domains	659:705	The base of the "V" is formed by two disulfide-linked dimeric N-terminal domains.					
25903135	5	77	with	coil	786:789	arg1	dimer					821:825	a C-terminal β-propeller dimer	796:825	a C-terminal β-propeller dimer at the tips	796:837	Each of the two V legs consists of a parallel dimeric disulfide-linked coiled coil with a C-terminal β-propeller dimer at the tips.					
25903135	3	78	theme	analytical	458:467	arg1	ultracentrifugation					469:487	analytical ultracentrifugation	458:487	analytical ultracentrifugation	458:487	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	3	79	theme	disulfide-linked	557:572	arg1	tetramers					574:582	disulfide-linked tetramers	557:582	disulfide-linked tetramers	557:582	Using a combined approach of x-ray crystallography, solution scattering, analytical ultracentrifugation, and electron microscopy we determined that full-length Olfm1 forms disulfide-linked tetramers with a distinctive V-shaped architecture.					
25903135	1	80	theme	nervous	249:255	arg1	development					264:274	nervous system development	249:274	nervous system development	249:274	Olfactomedin-1 (Olfm1; also known as noelin and pancortin) is a member of the olfactomedin domain-containing superfamily and a highly expressed neuronal glycoprotein important for nervous system development.