PubMed:8724132
Annnotations
sentences
{"project":"sentences","denotations":[{"id":"TextSentencer_T1","span":{"begin":0,"end":145},"obj":"Sentence"},{"id":"TextSentencer_T2","span":{"begin":146,"end":286},"obj":"Sentence"},{"id":"TextSentencer_T3","span":{"begin":287,"end":375},"obj":"Sentence"},{"id":"TextSentencer_T4","span":{"begin":376,"end":621},"obj":"Sentence"},{"id":"TextSentencer_T5","span":{"begin":622,"end":768},"obj":"Sentence"},{"id":"TextSentencer_T6","span":{"begin":769,"end":899},"obj":"Sentence"},{"id":"TextSentencer_T7","span":{"begin":900,"end":994},"obj":"Sentence"},{"id":"TextSentencer_T8","span":{"begin":995,"end":1147},"obj":"Sentence"},{"id":"TextSentencer_T9","span":{"begin":1148,"end":1253},"obj":"Sentence"},{"id":"TextSentencer_T10","span":{"begin":1254,"end":1478},"obj":"Sentence"},{"id":"TextSentencer_T11","span":{"begin":1479,"end":1586},"obj":"Sentence"},{"id":"TextSentencer_T12","span":{"begin":1587,"end":1676},"obj":"Sentence"},{"id":"TextSentencer_T13","span":{"begin":1677,"end":1900},"obj":"Sentence"},{"id":"TextSentencer_T14","span":{"begin":1901,"end":2014},"obj":"Sentence"},{"id":"TextSentencer_T15","span":{"begin":2015,"end":2223},"obj":"Sentence"},{"id":"T1","span":{"begin":0,"end":145},"obj":"Sentence"},{"id":"T2","span":{"begin":146,"end":286},"obj":"Sentence"},{"id":"T3","span":{"begin":287,"end":375},"obj":"Sentence"},{"id":"T4","span":{"begin":376,"end":621},"obj":"Sentence"},{"id":"T5","span":{"begin":622,"end":768},"obj":"Sentence"},{"id":"T6","span":{"begin":769,"end":899},"obj":"Sentence"},{"id":"T7","span":{"begin":900,"end":994},"obj":"Sentence"},{"id":"T8","span":{"begin":995,"end":1147},"obj":"Sentence"},{"id":"T9","span":{"begin":1148,"end":1253},"obj":"Sentence"},{"id":"T10","span":{"begin":1254,"end":1478},"obj":"Sentence"},{"id":"T11","span":{"begin":1479,"end":1586},"obj":"Sentence"},{"id":"T12","span":{"begin":1587,"end":1676},"obj":"Sentence"},{"id":"T13","span":{"begin":1677,"end":1900},"obj":"Sentence"},{"id":"T14","span":{"begin":1901,"end":2014},"obj":"Sentence"},{"id":"T15","span":{"begin":2015,"end":2223},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
GlycoBiology-FMA
{"project":"GlycoBiology-FMA","denotations":[{"id":"_T1","span":{"begin":235,"end":257},"obj":"FMAID:196780"},{"id":"_T2","span":{"begin":235,"end":257},"obj":"FMAID:82786"},{"id":"_T3","span":{"begin":679,"end":687},"obj":"FMAID:165447"},{"id":"_T4","span":{"begin":679,"end":687},"obj":"FMAID:67257"},{"id":"_T5","span":{"begin":734,"end":738},"obj":"FMAID:165676"},{"id":"_T6","span":{"begin":1165,"end":1171},"obj":"FMAID:178661"},{"id":"_T7","span":{"begin":1280,"end":1286},"obj":"FMAID:178661"},{"id":"_T8","span":{"begin":1681,"end":1693},"obj":"FMAID:197276"},{"id":"_T9","span":{"begin":1681,"end":1693},"obj":"FMAID:82737"},{"id":"_T10","span":{"begin":2162,"end":2173},"obj":"FMAID:164989"},{"id":"_T11","span":{"begin":2162,"end":2173},"obj":"FMAID:66835"},{"id":"_T12","span":{"begin":2162,"end":2173},"obj":"FMAID:165187"},{"id":"_T13","span":{"begin":2162,"end":2188},"obj":"FMAID:199171"}],"namespaces":[{"prefix":"FMAID","uri":"http://purl.org/sig/ont/fma/fma"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
uniprot-human
{"project":"uniprot-human","denotations":[{"id":"T1","span":{"begin":188,"end":192},"obj":"http://www.uniprot.org/uniprot/Q9NY76"},{"id":"T2","span":{"begin":309,"end":313},"obj":"http://www.uniprot.org/uniprot/Q9NY76"},{"id":"T3","span":{"begin":1767,"end":1771},"obj":"http://www.uniprot.org/uniprot/Q9NY76"},{"id":"T4","span":{"begin":496,"end":499},"obj":"http://www.uniprot.org/uniprot/P20132"},{"id":"T5","span":{"begin":1947,"end":1950},"obj":"http://www.uniprot.org/uniprot/P20132"},{"id":"T6","span":{"begin":2085,"end":2088},"obj":"http://www.uniprot.org/uniprot/P20132"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
uniprot-mouse
{"project":"uniprot-mouse","denotations":[{"id":"T1","span":{"begin":188,"end":192},"obj":"http://www.uniprot.org/uniprot/Q8JZM8"},{"id":"T2","span":{"begin":309,"end":313},"obj":"http://www.uniprot.org/uniprot/Q8JZM8"},{"id":"T3","span":{"begin":1767,"end":1771},"obj":"http://www.uniprot.org/uniprot/Q8JZM8"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
GlycoBiology-NCBITAXON
{"project":"GlycoBiology-NCBITAXON","denotations":[{"id":"T1","span":{"begin":177,"end":186},"obj":"http://purl.bioontology.org/ontology/STY/T192"},{"id":"T2","span":{"begin":846,"end":854},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/604139"},{"id":"T3","span":{"begin":1260,"end":1268},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/604139"},{"id":"T4","span":{"begin":1994,"end":2002},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/604139"},{"id":"T5","span":{"begin":2102,"end":2110},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/604139"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
GO-BP
{"project":"GO-BP","denotations":[{"id":"T1","span":{"begin":387,"end":403},"obj":"http://purl.obolibrary.org/obo/GO_0051099"},{"id":"T2","span":{"begin":1122,"end":1138},"obj":"http://purl.obolibrary.org/obo/GO_0051099"},{"id":"T3","span":{"begin":1220,"end":1236},"obj":"http://purl.obolibrary.org/obo/GO_0051099"},{"id":"T4","span":{"begin":1659,"end":1675},"obj":"http://purl.obolibrary.org/obo/GO_0051099"},{"id":"T5","span":{"begin":1815,"end":1831},"obj":"http://purl.obolibrary.org/obo/GO_0051099"},{"id":"T6","span":{"begin":2054,"end":2070},"obj":"http://purl.obolibrary.org/obo/GO_0051099"},{"id":"T7","span":{"begin":1754,"end":1763},"obj":"http://purl.obolibrary.org/obo/GO_0007586"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
GO-MF
{"project":"GO-MF","denotations":[{"id":"T1","span":{"begin":0,"end":12},"obj":"http://purl.obolibrary.org/obo/GO_0097617"},{"id":"T2","span":{"begin":878,"end":890},"obj":"http://purl.obolibrary.org/obo/GO_0097617"},{"id":"T3","span":{"begin":1200,"end":1208},"obj":"http://purl.obolibrary.org/obo/GO_0097617"},{"id":"T4","span":{"begin":17,"end":23},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T5","span":{"begin":380,"end":386},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T6","span":{"begin":447,"end":453},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T7","span":{"begin":625,"end":631},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T8","span":{"begin":1213,"end":1219},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T9","span":{"begin":1808,"end":1814},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T10","span":{"begin":1884,"end":1890},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T11","span":{"begin":2047,"end":2053},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T12","span":{"begin":197,"end":201},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T13","span":{"begin":387,"end":394},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T14","span":{"begin":632,"end":639},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T15","span":{"begin":910,"end":917},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T16","span":{"begin":1122,"end":1129},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T17","span":{"begin":1220,"end":1227},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T18","span":{"begin":1494,"end":1501},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T19","span":{"begin":1659,"end":1666},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T20","span":{"begin":1815,"end":1822},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T21","span":{"begin":1851,"end":1858},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T22","span":{"begin":1911,"end":1918},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T23","span":{"begin":2054,"end":2061},"obj":"http://purl.obolibrary.org/obo/GO_0070026"},{"id":"T24","span":{"begin":197,"end":201},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T25","span":{"begin":387,"end":394},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T26","span":{"begin":632,"end":639},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T27","span":{"begin":910,"end":917},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T28","span":{"begin":1122,"end":1129},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T29","span":{"begin":1220,"end":1227},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T30","span":{"begin":1494,"end":1501},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T31","span":{"begin":1659,"end":1666},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T32","span":{"begin":1815,"end":1822},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T33","span":{"begin":1851,"end":1858},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T34","span":{"begin":1911,"end":1918},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T35","span":{"begin":2054,"end":2061},"obj":"http://purl.obolibrary.org/obo/GO_0003680"},{"id":"T36","span":{"begin":197,"end":201},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T37","span":{"begin":387,"end":394},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T38","span":{"begin":632,"end":639},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T39","span":{"begin":910,"end":917},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T40","span":{"begin":1122,"end":1129},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T41","span":{"begin":1220,"end":1227},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T42","span":{"begin":1494,"end":1501},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T43","span":{"begin":1659,"end":1666},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T44","span":{"begin":1815,"end":1822},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T45","span":{"begin":1851,"end":1858},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T46","span":{"begin":1911,"end":1918},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T47","span":{"begin":2054,"end":2061},"obj":"http://purl.obolibrary.org/obo/GO_0017091"},{"id":"T48","span":{"begin":197,"end":201},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T49","span":{"begin":387,"end":394},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T50","span":{"begin":632,"end":639},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T51","span":{"begin":910,"end":917},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T52","span":{"begin":1122,"end":1129},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T53","span":{"begin":1220,"end":1227},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T54","span":{"begin":1494,"end":1501},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T55","span":{"begin":1659,"end":1666},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T56","span":{"begin":1815,"end":1822},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T57","span":{"begin":1851,"end":1858},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T58","span":{"begin":1911,"end":1918},"obj":"http://purl.obolibrary.org/obo/GO_0005488"},{"id":"T59","span":{"begin":2054,"end":2061},"obj":"http://purl.obolibrary.org/obo/GO_0005488"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
GO-CC
{"project":"GO-CC","denotations":[{"id":"T1","span":{"begin":2162,"end":2173},"obj":"http://purl.obolibrary.org/obo/GO_0005737"},{"id":"T2","span":{"begin":2175,"end":2188},"obj":"http://purl.obolibrary.org/obo/GO_0016021"},{"id":"T3","span":{"begin":2175,"end":2188},"obj":"http://purl.obolibrary.org/obo/GO_0044214"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
UBERON-AE
{"project":"UBERON-AE","denotations":[{"id":"T1","span":{"begin":734,"end":738},"obj":"http://purl.obolibrary.org/obo/UBERON_0001913"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
GlyCosmos600-FMA
{"project":"GlyCosmos600-FMA","denotations":[{"id":"T1","span":{"begin":679,"end":687},"obj":"Body_part"},{"id":"T2","span":{"begin":734,"end":738},"obj":"Body_part"},{"id":"T3","span":{"begin":1681,"end":1693},"obj":"Body_part"},{"id":"T4","span":{"begin":2162,"end":2173},"obj":"Body_part"}],"attributes":[{"id":"A1","pred":"fma_id","subj":"T1","obj":"http://purl.org/sig/ont/fma/fma67257"},{"id":"A2","pred":"fma_id","subj":"T2","obj":"http://purl.org/sig/ont/fma/fma62100"},{"id":"A3","pred":"fma_id","subj":"T3","obj":"http://purl.org/sig/ont/fma/fma82737"},{"id":"A4","pred":"fma_id","subj":"T4","obj":"http://purl.org/sig/ont/fma/fma66835"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
GlyCosmos600-GlycoProteins
{"project":"GlyCosmos600-GlycoProteins","denotations":[{"id":"PD-GlycoProteins-B_T1","span":{"begin":65,"end":92},"obj":"http://purl.uniprot.org/uniprot/P07307"},{"id":"PD-GlycoProteins-B_T2","span":{"begin":65,"end":92},"obj":"https://acgg.asia/db/gpdb/id/GPDB0003511"},{"id":"PD-GlycoProteins-B_T3","span":{"begin":65,"end":92},"obj":"http://purl.uniprot.org/uniprot/P07306"},{"id":"PD-GlycoProteins-B_T4","span":{"begin":65,"end":92},"obj":"https://acgg.asia/db/gpdb/id/GPDB0001846"},{"id":"PD-GlycoProteins-B_T5","span":{"begin":65,"end":92},"obj":"https://acgg.asia/db/gpdb/id/GPDB0003577"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
pubmed-enju-pas
{"project":"pubmed-enju-pas","denotations":[{"id":"EnjuParser_T0","span":{"begin":0,"end":12},"obj":"NN"},{"id":"EnjuParser_T1","span":{"begin":13,"end":16},"obj":"CC"},{"id":"EnjuParser_T2","span":{"begin":17,"end":23},"obj":"NN"},{"id":"EnjuParser_T3","span":{"begin":24,"end":32},"obj":"NN"},{"id":"EnjuParser_T4","span":{"begin":33,"end":35},"obj":"IN"},{"id":"EnjuParser_T5","span":{"begin":36,"end":39},"obj":"DT"},{"id":"EnjuParser_T6","span":{"begin":40,"end":45},"obj":"JJ"},{"id":"EnjuParser_T7","span":{"begin":46,"end":53},"obj":"NN"},{"id":"EnjuParser_T8","span":{"begin":54,"end":56},"obj":"IN"},{"id":"EnjuParser_T9","span":{"begin":57,"end":60},"obj":"DT"},{"id":"EnjuParser_T10","span":{"begin":61,"end":64},"obj":"NN"},{"id":"EnjuParser_T11","span":{"begin":65,"end":83},"obj":"NN"},{"id":"EnjuParser_T12","span":{"begin":84,"end":92},"obj":"NN"},{"id":"EnjuParser_T13","span":{"begin":93,"end":98},"obj":"IN"},{"id":"EnjuParser_T14","span":{"begin":99,"end":109},"obj":"VBG"},{"id":"EnjuParser_T15","span":{"begin":110,"end":124},"obj":"NN"},{"id":"EnjuParser_T16","span":{"begin":125,"end":128},"obj":"NN"},{"id":"EnjuParser_T17","span":{"begin":129,"end":144},"obj":"NN"},{"id":"EnjuParser_T18","span":{"begin":146,"end":149},"obj":"NN"},{"id":"EnjuParser_T19","span":{"begin":150,"end":157},"obj":"JJ"},{"id":"EnjuParser_T20","span":{"begin":158,"end":176},"obj":"NN"},{"id":"EnjuParser_T21","span":{"begin":177,"end":186},"obj":"NNS"},{"id":"EnjuParser_T22","span":{"begin":187,"end":188},"obj":"-LRB-"},{"id":"EnjuParser_T23","span":{"begin":188,"end":195},"obj":"NNS"},{"id":"EnjuParser_T24","span":{"begin":195,"end":196},"obj":"-RRB-"},{"id":"EnjuParser_T25","span":{"begin":197,"end":201},"obj":"VBP"},{"id":"EnjuParser_T26","span":{"begin":202,"end":210},"obj":"JJ"},{"id":"EnjuParser_T27","span":{"begin":211,"end":220},"obj":"VBN"},{"id":"EnjuParser_T28","span":{"begin":221,"end":231},"obj":"NN"},{"id":"EnjuParser_T29","span":{"begin":232,"end":234},"obj":"CC"},{"id":"EnjuParser_T30","span":{"begin":235,"end":257},"obj":"NN"},{"id":"EnjuParser_T31","span":{"begin":258,"end":266},"obj":"NNS"},{"id":"EnjuParser_T32","span":{"begin":267,"end":271},"obj":"IN"},{"id":"EnjuParser_T33","span":{"begin":272,"end":276},"obj":"JJ"},{"id":"EnjuParser_T34","span":{"begin":277,"end":285},"obj":"NN"},{"id":"EnjuParser_T35","span":{"begin":287,"end":290},"obj":"DT"},{"id":"EnjuParser_T36","span":{"begin":291,"end":308},"obj":"JJ"},{"id":"EnjuParser_T37","span":{"begin":309,"end":315},"obj":"NN"},{"id":"EnjuParser_T38","span":{"begin":316,"end":324},"obj":"VBZ"},{"id":"EnjuParser_T39","span":{"begin":325,"end":327},"obj":"IN"},{"id":"EnjuParser_T40","span":{"begin":328,"end":333},"obj":"CD"},{"id":"EnjuParser_T41","span":{"begin":334,"end":342},"obj":"NNS"},{"id":"EnjuParser_T42","span":{"begin":343,"end":353},"obj":"VBN"},{"id":"EnjuParser_T43","span":{"begin":354,"end":358},"obj":"NN"},{"id":"EnjuParser_T44","span":{"begin":358,"end":359},"obj":"-COMMA-"},{"id":"EnjuParser_T45","span":{"begin":360,"end":364},"obj":"NN"},{"id":"EnjuParser_T46","span":{"begin":364,"end":365},"obj":"-COMMA-"},{"id":"EnjuParser_T47","span":{"begin":366,"end":369},"obj":"CC"},{"id":"EnjuParser_T48","span":{"begin":370,"end":374},"obj":"NN"},{"id":"EnjuParser_T49","span":{"begin":376,"end":379},"obj":"DT"},{"id":"EnjuParser_T50","span":{"begin":380,"end":394},"obj":"JJ"},{"id":"EnjuParser_T51","span":{"begin":395,"end":403},"obj":"NN"},{"id":"EnjuParser_T52","span":{"begin":404,"end":406},"obj":"IN"},{"id":"EnjuParser_T53","span":{"begin":407,"end":417},"obj":"JJ"},{"id":"EnjuParser_T54","span":{"begin":418,"end":426},"obj":"NNS"},{"id":"EnjuParser_T55","span":{"begin":427,"end":430},"obj":"VBD"},{"id":"EnjuParser_T56","span":{"begin":431,"end":443},"obj":"VBN"},{"id":"EnjuParser_T57","span":{"begin":444,"end":446},"obj":"IN"},{"id":"EnjuParser_T58","span":{"begin":447,"end":453},"obj":"NN"},{"id":"EnjuParser_T59","span":{"begin":454,"end":462},"obj":"NN"},{"id":"EnjuParser_T60","span":{"begin":462,"end":463},"obj":"-COMMA-"},{"id":"EnjuParser_T61","span":{"begin":464,"end":469},"obj":"IN"},{"id":"EnjuParser_T62","span":{"begin":470,"end":480},"obj":"NN"},{"id":"EnjuParser_T63","span":{"begin":481,"end":483},"obj":"IN"},{"id":"EnjuParser_T64","span":{"begin":484,"end":492},"obj":"NNS"},{"id":"EnjuParser_T65","span":{"begin":493,"end":495},"obj":"IN"},{"id":"EnjuParser_T66","span":{"begin":496,"end":504},"obj":"NN"},{"id":"EnjuParser_T67","span":{"begin":505,"end":510},"obj":"IN"},{"id":"EnjuParser_T68","span":{"begin":511,"end":522},"obj":"JJ"},{"id":"EnjuParser_T69","span":{"begin":523,"end":533},"obj":"NNS"},{"id":"EnjuParser_T70","span":{"begin":533,"end":534},"obj":"-COMMA-"},{"id":"EnjuParser_T71","span":{"begin":535,"end":550},"obj":"NN"},{"id":"EnjuParser_T72","span":{"begin":551,"end":553},"obj":"TO"},{"id":"EnjuParser_T73","span":{"begin":554,"end":568},"obj":"NN"},{"id":"EnjuParser_T74","span":{"begin":568,"end":569},"obj":"-COMMA-"},{"id":"EnjuParser_T75","span":{"begin":570,"end":573},"obj":"CC"},{"id":"EnjuParser_T76","span":{"begin":574,"end":584},"obj":"NN"},{"id":"EnjuParser_T77","span":{"begin":585,"end":589},"obj":"IN"},{"id":"EnjuParser_T78","span":{"begin":590,"end":613},"obj":"NN"},{"id":"EnjuParser_T79","span":{"begin":614,"end":615},"obj":"-LRB-"},{"id":"EnjuParser_T80","span":{"begin":615,"end":619},"obj":"NN"},{"id":"EnjuParser_T81","span":{"begin":619,"end":620},"obj":"-RRB-"},{"id":"EnjuParser_T82","span":{"begin":622,"end":624},"obj":"DT"},{"id":"EnjuParser_T83","span":{"begin":625,"end":639},"obj":"JJ"},{"id":"EnjuParser_T84","span":{"begin":640,"end":642},"obj":"TO"},{"id":"EnjuParser_T85","span":{"begin":643,"end":646},"obj":"DT"},{"id":"EnjuParser_T86","span":{"begin":647,"end":655},"obj":"NNS"},{"id":"EnjuParser_T87","span":{"begin":656,"end":661},"obj":"MD"},{"id":"EnjuParser_T88","span":{"begin":662,"end":664},"obj":"VB"},{"id":"EnjuParser_T89","span":{"begin":665,"end":673},"obj":"VBN"},{"id":"EnjuParser_T90","span":{"begin":674,"end":678},"obj":"WRB"},{"id":"EnjuParser_T91","span":{"begin":679,"end":687},"obj":"NNS"},{"id":"EnjuParser_T92","span":{"begin":688,"end":692},"obj":"JJ"},{"id":"EnjuParser_T93","span":{"begin":693,"end":695},"obj":"IN"},{"id":"EnjuParser_T94","span":{"begin":696,"end":699},"obj":"NN"},{"id":"EnjuParser_T95","span":{"begin":699,"end":700},"obj":"-COMMA-"},{"id":"EnjuParser_T96","span":{"begin":701,"end":707},"obj":"NN"},{"id":"EnjuParser_T97","span":{"begin":707,"end":708},"obj":"-COMMA-"},{"id":"EnjuParser_T98","span":{"begin":709,"end":716},"obj":"NN"},{"id":"EnjuParser_T99","span":{"begin":716,"end":717},"obj":"-COMMA-"},{"id":"EnjuParser_T100","span":{"begin":718,"end":720},"obj":"CC"},{"id":"EnjuParser_T101","span":{"begin":721,"end":729},"obj":"JJ"},{"id":"EnjuParser_T102","span":{"begin":730,"end":733},"obj":"JJ"},{"id":"EnjuParser_T103","span":{"begin":734,"end":738},"obj":"NN"},{"id":"EnjuParser_T104","span":{"begin":739,"end":743},"obj":"VBD"},{"id":"EnjuParser_T105","span":{"begin":744,"end":748},"obj":"VBN"},{"id":"EnjuParser_T106","span":{"begin":749,"end":751},"obj":"IN"},{"id":"EnjuParser_T107","span":{"begin":752,"end":760},"obj":"VBG"},{"id":"EnjuParser_T108","span":{"begin":761,"end":767},"obj":"NNS"},{"id":"EnjuParser_T109","span":{"begin":769,"end":776},"obj":"RB"},{"id":"EnjuParser_T110","span":{"begin":776,"end":777},"obj":"-COMMA-"},{"id":"EnjuParser_T111","span":{"begin":778,"end":788},"obj":"JJ"},{"id":"EnjuParser_T112","span":{"begin":789,"end":799},"obj":"NN"},{"id":"EnjuParser_T113","span":{"begin":800,"end":802},"obj":"IN"},{"id":"EnjuParser_T114","span":{"begin":803,"end":814},"obj":"JJ"},{"id":"EnjuParser_T115","span":{"begin":815,"end":829},"obj":"NN"},{"id":"EnjuParser_T116","span":{"begin":830,"end":835},"obj":"NNS"},{"id":"EnjuParser_T117","span":{"begin":836,"end":840},"obj":"IN"},{"id":"EnjuParser_T118","span":{"begin":841,"end":845},"obj":"DT"},{"id":"EnjuParser_T119","span":{"begin":846,"end":854},"obj":"JJ"},{"id":"EnjuParser_T120","span":{"begin":855,"end":865},"obj":"NNS"},{"id":"EnjuParser_T121","span":{"begin":866,"end":874},"obj":"VBD"},{"id":"EnjuParser_T122","span":{"begin":875,"end":877},"obj":"IN"},{"id":"EnjuParser_T123","span":{"begin":878,"end":890},"obj":"NN"},{"id":"EnjuParser_T124","span":{"begin":891,"end":893},"obj":"IN"},{"id":"EnjuParser_T125","span":{"begin":894,"end":898},"obj":"NN"},{"id":"EnjuParser_T126","span":{"begin":900,"end":917},"obj":"JJ"},{"id":"EnjuParser_T127","span":{"begin":918,"end":920},"obj":"TO"},{"id":"EnjuParser_T128","span":{"begin":921,"end":925},"obj":"NN"},{"id":"EnjuParser_T129","span":{"begin":926,"end":928},"obj":"CC"},{"id":"EnjuParser_T130","span":{"begin":929,"end":933},"obj":"NN"},{"id":"EnjuParser_T131","span":{"begin":934,"end":937},"obj":"VBD"},{"id":"EnjuParser_T132","span":{"begin":938,"end":944},"obj":"JJR"},{"id":"EnjuParser_T133","span":{"begin":945,"end":948},"obj":"CC"},{"id":"EnjuParser_T134","span":{"begin":949,"end":954},"obj":"MD"},{"id":"EnjuParser_T135","span":{"begin":955,"end":957},"obj":"VB"},{"id":"EnjuParser_T136","span":{"begin":958,"end":966},"obj":"VBN"},{"id":"EnjuParser_T137","span":{"begin":967,"end":971},"obj":"RB"},{"id":"EnjuParser_T138","span":{"begin":972,"end":977},"obj":"IN"},{"id":"EnjuParser_T139","span":{"begin":978,"end":984},"obj":"JJR"},{"id":"EnjuParser_T140","span":{"begin":985,"end":993},"obj":"NN"},{"id":"EnjuParser_T141","span":{"begin":995,"end":1004},"obj":"RB"},{"id":"EnjuParser_T142","span":{"begin":1004,"end":1005},"obj":"-COMMA-"},{"id":"EnjuParser_T143","span":{"begin":1006,"end":1012},"obj":"JJ"},{"id":"EnjuParser_T144","span":{"begin":1013,"end":1016},"obj":"NN"},{"id":"EnjuParser_T145","span":{"begin":1017,"end":1019},"obj":"IN"},{"id":"EnjuParser_T146","span":{"begin":1020,"end":1030},"obj":"NNS"},{"id":"EnjuParser_T147","span":{"begin":1031,"end":1035},"obj":"JJ"},{"id":"EnjuParser_T148","span":{"begin":1036,"end":1038},"obj":"IN"},{"id":"EnjuParser_T149","span":{"begin":1039,"end":1044},"obj":"CD"},{"id":"EnjuParser_T150","span":{"begin":1045,"end":1047},"obj":"CD"},{"id":"EnjuParser_T151","span":{"begin":1047,"end":1048},"obj":"-COMMA-"},{"id":"EnjuParser_T152","span":{"begin":1049,"end":1056},"obj":"NNP"},{"id":"EnjuParser_T153","span":{"begin":1057,"end":1061},"obj":"NN"},{"id":"EnjuParser_T154","span":{"begin":1061,"end":1062},"obj":"-COMMA-"},{"id":"EnjuParser_T155","span":{"begin":1063,"end":1065},"obj":"CC"},{"id":"EnjuParser_T156","span":{"begin":1066,"end":1072},"obj":"NN"},{"id":"EnjuParser_T157","span":{"begin":1073,"end":1078},"obj":"NN"},{"id":"EnjuParser_T158","span":{"begin":1079,"end":1081},"obj":"IN"},{"id":"EnjuParser_T159","span":{"begin":1082,"end":1090},"obj":"VBG"},{"id":"EnjuParser_T160","span":{"begin":1091,"end":1097},"obj":"NNS"},{"id":"EnjuParser_T161","span":{"begin":1098,"end":1102},"obj":"RB"},{"id":"EnjuParser_T162","span":{"begin":1103,"end":1112},"obj":"VBD"},{"id":"EnjuParser_T163","span":{"begin":1113,"end":1116},"obj":"DT"},{"id":"EnjuParser_T164","span":{"begin":1117,"end":1129},"obj":"JJ"},{"id":"EnjuParser_T165","span":{"begin":1130,"end":1138},"obj":"NN"},{"id":"EnjuParser_T166","span":{"begin":1139,"end":1141},"obj":"IN"},{"id":"EnjuParser_T167","span":{"begin":1142,"end":1146},"obj":"NN"},{"id":"EnjuParser_T168","span":{"begin":1148,"end":1153},"obj":"JJ"},{"id":"EnjuParser_T169","span":{"begin":1154,"end":1164},"obj":"NNS"},{"id":"EnjuParser_T170","span":{"begin":1165,"end":1171},"obj":"VBN"},{"id":"EnjuParser_T171","span":{"begin":1172,"end":1175},"obj":"VBD"},{"id":"EnjuParser_T172","span":{"begin":1176,"end":1179},"obj":"RB"},{"id":"EnjuParser_T173","span":{"begin":1180,"end":1184},"obj":"VB"},{"id":"EnjuParser_T174","span":{"begin":1185,"end":1188},"obj":"DT"},{"id":"EnjuParser_T175","span":{"begin":1189,"end":1196},"obj":"NN"},{"id":"EnjuParser_T176","span":{"begin":1197,"end":1199},"obj":"TO"},{"id":"EnjuParser_T177","span":{"begin":1200,"end":1208},"obj":"VB"},{"id":"EnjuParser_T178","span":{"begin":1209,"end":1212},"obj":"DT"},{"id":"EnjuParser_T179","span":{"begin":1213,"end":1227},"obj":"JJ"},{"id":"EnjuParser_T180","span":{"begin":1228,"end":1236},"obj":"NN"},{"id":"EnjuParser_T181","span":{"begin":1237,"end":1239},"obj":"IN"},{"id":"EnjuParser_T182","span":{"begin":1240,"end":1243},"obj":"NN"},{"id":"EnjuParser_T183","span":{"begin":1244,"end":1252},"obj":"NNS"},{"id":"EnjuParser_T184","span":{"begin":1254,"end":1259},"obj":"IN"},{"id":"EnjuParser_T185","span":{"begin":1260,"end":1268},"obj":"JJ"},{"id":"EnjuParser_T186","span":{"begin":1269,"end":1279},"obj":"NNS"},{"id":"EnjuParser_T187","span":{"begin":1280,"end":1286},"obj":"VBN"},{"id":"EnjuParser_T188","span":{"begin":1286,"end":1287},"obj":"-COMMA-"},{"id":"EnjuParser_T189","span":{"begin":1288,"end":1293},"obj":"NNP"},{"id":"EnjuParser_T190","span":{"begin":1294,"end":1296},"obj":"CD"},{"id":"EnjuParser_T191","span":{"begin":1296,"end":1297},"obj":"-COMMA-"},{"id":"EnjuParser_T192","span":{"begin":1298,"end":1303},"obj":"CD"},{"id":"EnjuParser_T193","span":{"begin":1304,"end":1306},"obj":"CD"},{"id":"EnjuParser_T194","span":{"begin":1306,"end":1307},"obj":"-COMMA-"},{"id":"EnjuParser_T195","span":{"begin":1308,"end":1314},"obj":"NNP"},{"id":"EnjuParser_T196","span":{"begin":1315,"end":1317},"obj":"NNP"},{"id":"EnjuParser_T197","span":{"begin":1317,"end":1318},"obj":"-COMMA-"},{"id":"EnjuParser_T198","span":{"begin":1319,"end":1326},"obj":"NNP"},{"id":"EnjuParser_T199","span":{"begin":1327,"end":1331},"obj":"NN"},{"id":"EnjuParser_T200","span":{"begin":1331,"end":1332},"obj":"-COMMA-"},{"id":"EnjuParser_T201","span":{"begin":1333,"end":1336},"obj":"CC"},{"id":"EnjuParser_T202","span":{"begin":1337,"end":1343},"obj":"NN"},{"id":"EnjuParser_T203","span":{"begin":1344,"end":1349},"obj":"NN"},{"id":"EnjuParser_T204","span":{"begin":1350,"end":1354},"obj":"VBD"},{"id":"EnjuParser_T205","span":{"begin":1355,"end":1359},"obj":"RBR"},{"id":"EnjuParser_T206","span":{"begin":1360,"end":1369},"obj":"JJ"},{"id":"EnjuParser_T207","span":{"begin":1370,"end":1374},"obj":"IN"},{"id":"EnjuParser_T208","span":{"begin":1375,"end":1380},"obj":"CD"},{"id":"EnjuParser_T209","span":{"begin":1381,"end":1383},"obj":"CD"},{"id":"EnjuParser_T210","span":{"begin":1383,"end":1384},"obj":"-COMMA-"},{"id":"EnjuParser_T211","span":{"begin":1385,"end":1390},"obj":"CD"},{"id":"EnjuParser_T212","span":{"begin":1391,"end":1393},"obj":"CD"},{"id":"EnjuParser_T213","span":{"begin":1393,"end":1394},"obj":"-COMMA-"},{"id":"EnjuParser_T214","span":{"begin":1395,"end":1400},"obj":"CD"},{"id":"EnjuParser_T215","span":{"begin":1401,"end":1403},"obj":"CD"},{"id":"EnjuParser_T216","span":{"begin":1403,"end":1404},"obj":"-COMMA-"},{"id":"EnjuParser_T217","span":{"begin":1405,"end":1407},"obj":"CC"},{"id":"EnjuParser_T218","span":{"begin":1408,"end":1412},"obj":"NNP"},{"id":"EnjuParser_T219","span":{"begin":1413,"end":1415},"obj":"CD"},{"id":"EnjuParser_T220","span":{"begin":1415,"end":1416},"obj":"-COMMA-"},{"id":"EnjuParser_T221","span":{"begin":1417,"end":1424},"obj":"IN"},{"id":"EnjuParser_T222","span":{"begin":1425,"end":1429},"obj":"NN"},{"id":"EnjuParser_T223","span":{"begin":1429,"end":1430},"obj":"-COMMA-"},{"id":"EnjuParser_T224","span":{"begin":1431,"end":1440},"obj":"NN"},{"id":"EnjuParser_T225","span":{"begin":1440,"end":1441},"obj":"-COMMA-"},{"id":"EnjuParser_T226","span":{"begin":1442,"end":1444},"obj":"CC"},{"id":"EnjuParser_T227","span":{"begin":1445,"end":1460},"obj":"NN"},{"id":"EnjuParser_T228","span":{"begin":1461,"end":1467},"obj":"VBD"},{"id":"EnjuParser_T229","span":{"begin":1468,"end":1470},"obj":"DT"},{"id":"EnjuParser_T230","span":{"begin":1471,"end":1477},"obj":"NN"},{"id":"EnjuParser_T231","span":{"begin":1479,"end":1483},"obj":"JJ"},{"id":"EnjuParser_T232","span":{"begin":1484,"end":1493},"obj":"NN"},{"id":"EnjuParser_T233","span":{"begin":1494,"end":1501},"obj":"VBG"},{"id":"EnjuParser_T234","span":{"begin":1502,"end":1504},"obj":"TO"},{"id":"EnjuParser_T235","span":{"begin":1505,"end":1509},"obj":"NN"},{"id":"EnjuParser_T236","span":{"begin":1510,"end":1512},"obj":"CC"},{"id":"EnjuParser_T237","span":{"begin":1513,"end":1517},"obj":"NN"},{"id":"EnjuParser_T238","span":{"begin":1518,"end":1523},"obj":"MD"},{"id":"EnjuParser_T239","span":{"begin":1524,"end":1526},"obj":"VB"},{"id":"EnjuParser_T240","span":{"begin":1527,"end":1535},"obj":"VBN"},{"id":"EnjuParser_T241","span":{"begin":1536,"end":1540},"obj":"RB"},{"id":"EnjuParser_T242","span":{"begin":1541,"end":1545},"obj":"WRB"},{"id":"EnjuParser_T243","span":{"begin":1546,"end":1549},"obj":"DT"},{"id":"EnjuParser_T244","span":{"begin":1550,"end":1555},"obj":"CD"},{"id":"EnjuParser_T245","span":{"begin":1556,"end":1562},"obj":"NN"},{"id":"EnjuParser_T246","span":{"begin":1563,"end":1565},"obj":"CC"},{"id":"EnjuParser_T247","span":{"begin":1566,"end":1572},"obj":"NN"},{"id":"EnjuParser_T248","span":{"begin":1573,"end":1575},"obj":"NN"},{"id":"EnjuParser_T249","span":{"begin":1576,"end":1580},"obj":"VBD"},{"id":"EnjuParser_T250","span":{"begin":1581,"end":1585},"obj":"VBN"},{"id":"EnjuParser_T251","span":{"begin":1587,"end":1597},"obj":"NN"},{"id":"EnjuParser_T252","span":{"begin":1598,"end":1600},"obj":"IN"},{"id":"EnjuParser_T253","span":{"begin":1601,"end":1606},"obj":"NNS"},{"id":"EnjuParser_T254","span":{"begin":1607,"end":1611},"obj":"IN"},{"id":"EnjuParser_T255","span":{"begin":1612,"end":1626},"obj":"NN"},{"id":"EnjuParser_T256","span":{"begin":1627,"end":1633},"obj":"VBD"},{"id":"EnjuParser_T257","span":{"begin":1634,"end":1635},"obj":"DT"},{"id":"EnjuParser_T258","span":{"begin":1636,"end":1650},"obj":"JJ"},{"id":"EnjuParser_T259","span":{"begin":1651,"end":1655},"obj":"NN"},{"id":"EnjuParser_T260","span":{"begin":1656,"end":1658},"obj":"IN"},{"id":"EnjuParser_T261","span":{"begin":1659,"end":1666},"obj":"NN"},{"id":"EnjuParser_T262","span":{"begin":1667,"end":1675},"obj":"NN"},{"id":"EnjuParser_T263","span":{"begin":1677,"end":1680},"obj":"DT"},{"id":"EnjuParser_T264","span":{"begin":1681,"end":1693},"obj":"NN"},{"id":"EnjuParser_T265","span":{"begin":1694,"end":1705},"obj":"NN"},{"id":"EnjuParser_T266","span":{"begin":1706,"end":1712},"obj":"NN"},{"id":"EnjuParser_T267","span":{"begin":1713,"end":1714},"obj":"-LRB-"},{"id":"EnjuParser_T268","span":{"begin":1714,"end":1717},"obj":"NN"},{"id":"EnjuParser_T269","span":{"begin":1717,"end":1718},"obj":"-RRB-"},{"id":"EnjuParser_T270","span":{"begin":1719,"end":1721},"obj":"IN"},{"id":"EnjuParser_T271","span":{"begin":1722,"end":1726},"obj":"NN"},{"id":"EnjuParser_T272","span":{"begin":1726,"end":1727},"obj":"-COMMA-"},{"id":"EnjuParser_T273","span":{"begin":1728,"end":1736},"obj":"VBN"},{"id":"EnjuParser_T274","span":{"begin":1737,"end":1742},"obj":"IN"},{"id":"EnjuParser_T275","span":{"begin":1743,"end":1753},"obj":"NN"},{"id":"EnjuParser_T276","span":{"begin":1754,"end":1763},"obj":"NN"},{"id":"EnjuParser_T277","span":{"begin":1764,"end":1766},"obj":"IN"},{"id":"EnjuParser_T278","span":{"begin":1767,"end":1773},"obj":"NN"},{"id":"EnjuParser_T279","span":{"begin":1774,"end":1779},"obj":"VBD"},{"id":"EnjuParser_T280","span":{"begin":1780,"end":1782},"obj":"TO"},{"id":"EnjuParser_T281","span":{"begin":1783,"end":1797},"obj":"NN"},{"id":"EnjuParser_T282","span":{"begin":1797,"end":1798},"obj":"-COMMA-"},{"id":"EnjuParser_T283","span":{"begin":1799,"end":1807},"obj":"VBD"},{"id":"EnjuParser_T284","span":{"begin":1808,"end":1822},"obj":"JJ"},{"id":"EnjuParser_T285","span":{"begin":1823,"end":1831},"obj":"NN"},{"id":"EnjuParser_T286","span":{"begin":1832,"end":1834},"obj":"IN"},{"id":"EnjuParser_T287","span":{"begin":1835,"end":1843},"obj":"VBN"},{"id":"EnjuParser_T288","span":{"begin":1844,"end":1846},"obj":"IN"},{"id":"EnjuParser_T289","span":{"begin":1847,"end":1850},"obj":"PRP-DOLLAR-"},{"id":"EnjuParser_T290","span":{"begin":1851,"end":1858},"obj":"NN"},{"id":"EnjuParser_T291","span":{"begin":1859,"end":1861},"obj":"TO"},{"id":"EnjuParser_T292","span":{"begin":1862,"end":1876},"obj":"NN"},{"id":"EnjuParser_T293","span":{"begin":1877,"end":1880},"obj":"CC"},{"id":"EnjuParser_T294","span":{"begin":1881,"end":1883},"obj":"IN"},{"id":"EnjuParser_T295","span":{"begin":1884,"end":1890},"obj":"NN"},{"id":"EnjuParser_T296","span":{"begin":1891,"end":1899},"obj":"NN"},{"id":"EnjuParser_T297","span":{"begin":1901,"end":1910},"obj":"NN"},{"id":"EnjuParser_T298","span":{"begin":1911,"end":1918},"obj":"NN"},{"id":"EnjuParser_T299","span":{"begin":1919,"end":1921},"obj":"TO"},{"id":"EnjuParser_T300","span":{"begin":1922,"end":1936},"obj":"VBN"},{"id":"EnjuParser_T301","span":{"begin":1937,"end":1940},"obj":"NN"},{"id":"EnjuParser_T302","span":{"begin":1941,"end":1946},"obj":"IN"},{"id":"EnjuParser_T303","span":{"begin":1947,"end":1955},"obj":"NN"},{"id":"EnjuParser_T304","span":{"begin":1956,"end":1959},"obj":"VBD"},{"id":"EnjuParser_T305","span":{"begin":1960,"end":1964},"obj":"RB"},{"id":"EnjuParser_T306","span":{"begin":1965,"end":1974},"obj":"JJ"},{"id":"EnjuParser_T307","span":{"begin":1975,"end":1977},"obj":"IN"},{"id":"EnjuParser_T308","span":{"begin":1978,"end":1981},"obj":"DT"},{"id":"EnjuParser_T309","span":{"begin":1982,"end":1990},"obj":"NN"},{"id":"EnjuParser_T310","span":{"begin":1991,"end":1993},"obj":"IN"},{"id":"EnjuParser_T311","span":{"begin":1994,"end":2002},"obj":"JJ"},{"id":"EnjuParser_T312","span":{"begin":2003,"end":2013},"obj":"NNS"},{"id":"EnjuParser_T313","span":{"begin":2015,"end":2017},"obj":"PRP"},{"id":"EnjuParser_T314","span":{"begin":2018,"end":2026},"obj":"VBP"},{"id":"EnjuParser_T315","span":{"begin":2027,"end":2031},"obj":"IN"},{"id":"EnjuParser_T316","span":{"begin":2032,"end":2043},"obj":"NN"},{"id":"EnjuParser_T317","span":{"begin":2044,"end":2046},"obj":"IN"},{"id":"EnjuParser_T318","span":{"begin":2047,"end":2061},"obj":"JJ"},{"id":"EnjuParser_T319","span":{"begin":2062,"end":2070},"obj":"NN"},{"id":"EnjuParser_T320","span":{"begin":2071,"end":2073},"obj":"IN"},{"id":"EnjuParser_T321","span":{"begin":2074,"end":2078},"obj":"NN"},{"id":"EnjuParser_T322","span":{"begin":2079,"end":2084},"obj":"IN"},{"id":"EnjuParser_T323","span":{"begin":2085,"end":2093},"obj":"NN"},{"id":"EnjuParser_T324","span":{"begin":2094,"end":2096},"obj":"IN"},{"id":"EnjuParser_T325","span":{"begin":2097,"end":2101},"obj":"DT"},{"id":"EnjuParser_T326","span":{"begin":2102,"end":2110},"obj":"JJ"},{"id":"EnjuParser_T327","span":{"begin":2111,"end":2121},"obj":"NNS"},{"id":"EnjuParser_T328","span":{"begin":2122,"end":2124},"obj":"VBZ"},{"id":"EnjuParser_T329","span":{"begin":2125,"end":2128},"obj":"RB"},{"id":"EnjuParser_T330","span":{"begin":2129,"end":2138},"obj":"JJ"},{"id":"EnjuParser_T331","span":{"begin":2139,"end":2141},"obj":"IN"},{"id":"EnjuParser_T332","span":{"begin":2142,"end":2145},"obj":"DT"},{"id":"EnjuParser_T333","span":{"begin":2146,"end":2154},"obj":"NN"},{"id":"EnjuParser_T334","span":{"begin":2155,"end":2157},"obj":"IN"},{"id":"EnjuParser_T335","span":{"begin":2158,"end":2161},"obj":"DT"},{"id":"EnjuParser_T336","span":{"begin":2162,"end":2173},"obj":"JJ"},{"id":"EnjuParser_T337","span":{"begin":2173,"end":2174},"obj":"-COMMA-"},{"id":"EnjuParser_T338","span":{"begin":2175,"end":2188},"obj":"NN"},{"id":"EnjuParser_T339","span":{"begin":2188,"end":2189},"obj":"-COMMA-"},{"id":"EnjuParser_T340","span":{"begin":2190,"end":2192},"obj":"CC"},{"id":"EnjuParser_T341","span":{"begin":2193,"end":2198},"obj":"NN"},{"id":"EnjuParser_T342","span":{"begin":2199,"end":2206},"obj":"NNS"},{"id":"EnjuParser_T343","span":{"begin":2207,"end":2209},"obj":"IN"},{"id":"EnjuParser_T344","span":{"begin":2210,"end":2214},"obj":"DT"},{"id":"EnjuParser_T345","span":{"begin":2215,"end":2222},"obj":"NN"}],"relations":[{"id":"EnjuParser_R0","pred":"arg1Of","subj":"EnjuParser_T3","obj":"EnjuParser_T0"},{"id":"EnjuParser_R1","pred":"arg1Of","subj":"EnjuParser_T0","obj":"EnjuParser_T1"},{"id":"EnjuParser_R2","pred":"arg2Of","subj":"EnjuParser_T2","obj":"EnjuParser_T1"},{"id":"EnjuParser_R3","pred":"arg1Of","subj":"EnjuParser_T3","obj":"EnjuParser_T2"},{"id":"EnjuParser_R4","pred":"arg1Of","subj":"EnjuParser_T3","obj":"EnjuParser_T4"},{"id":"EnjuParser_R5","pred":"arg2Of","subj":"EnjuParser_T7","obj":"EnjuParser_T4"},{"id":"EnjuParser_R6","pred":"arg1Of","subj":"EnjuParser_T7","obj":"EnjuParser_T5"},{"id":"EnjuParser_R7","pred":"arg1Of","subj":"EnjuParser_T7","obj":"EnjuParser_T6"},{"id":"EnjuParser_R8","pred":"arg1Of","subj":"EnjuParser_T7","obj":"EnjuParser_T8"},{"id":"EnjuParser_R9","pred":"arg2Of","subj":"EnjuParser_T12","obj":"EnjuParser_T8"},{"id":"EnjuParser_R10","pred":"arg1Of","subj":"EnjuParser_T12","obj":"EnjuParser_T9"},{"id":"EnjuParser_R11","pred":"arg1Of","subj":"EnjuParser_T12","obj":"EnjuParser_T10"},{"id":"EnjuParser_R12","pred":"arg1Of","subj":"EnjuParser_T12","obj":"EnjuParser_T11"},{"id":"EnjuParser_R13","pred":"arg1Of","subj":"EnjuParser_T3","obj":"EnjuParser_T13"},{"id":"EnjuParser_R14","pred":"arg2Of","subj":"EnjuParser_T14","obj":"EnjuParser_T13"},{"id":"EnjuParser_R15","pred":"arg2Of","subj":"EnjuParser_T17","obj":"EnjuParser_T14"},{"id":"EnjuParser_R16","pred":"arg1Of","subj":"EnjuParser_T17","obj":"EnjuParser_T15"},{"id":"EnjuParser_R17","pred":"arg1Of","subj":"EnjuParser_T17","obj":"EnjuParser_T16"},{"id":"EnjuParser_R18","pred":"arg1Of","subj":"EnjuParser_T21","obj":"EnjuParser_T18"},{"id":"EnjuParser_R19","pred":"arg1Of","subj":"EnjuParser_T21","obj":"EnjuParser_T19"},{"id":"EnjuParser_R20","pred":"arg1Of","subj":"EnjuParser_T21","obj":"EnjuParser_T20"},{"id":"EnjuParser_R21","pred":"arg1Of","subj":"EnjuParser_T21","obj":"EnjuParser_T22"},{"id":"EnjuParser_R22","pred":"arg2Of","subj":"EnjuParser_T23","obj":"EnjuParser_T22"},{"id":"EnjuParser_R23","pred":"arg3Of","subj":"EnjuParser_T24","obj":"EnjuParser_T22"},{"id":"EnjuParser_R24","pred":"arg1Of","subj":"EnjuParser_T21","obj":"EnjuParser_T25"},{"id":"EnjuParser_R25","pred":"arg2Of","subj":"EnjuParser_T31","obj":"EnjuParser_T25"},{"id":"EnjuParser_R26","pred":"arg1Of","subj":"EnjuParser_T31","obj":"EnjuParser_T26"},{"id":"EnjuParser_R27","pred":"arg2Of","subj":"EnjuParser_T31","obj":"EnjuParser_T27"},{"id":"EnjuParser_R28","pred":"arg1Of","subj":"EnjuParser_T31","obj":"EnjuParser_T28"},{"id":"EnjuParser_R29","pred":"arg1Of","subj":"EnjuParser_T28","obj":"EnjuParser_T29"},{"id":"EnjuParser_R30","pred":"arg2Of","subj":"EnjuParser_T30","obj":"EnjuParser_T29"},{"id":"EnjuParser_R31","pred":"arg1Of","subj":"EnjuParser_T31","obj":"EnjuParser_T30"},{"id":"EnjuParser_R32","pred":"arg1Of","subj":"EnjuParser_T31","obj":"EnjuParser_T32"},{"id":"EnjuParser_R33","pred":"arg2Of","subj":"EnjuParser_T34","obj":"EnjuParser_T32"},{"id":"EnjuParser_R34","pred":"arg1Of","subj":"EnjuParser_T34","obj":"EnjuParser_T33"},{"id":"EnjuParser_R35","pred":"arg1Of","subj":"EnjuParser_T37","obj":"EnjuParser_T35"},{"id":"EnjuParser_R36","pred":"arg1Of","subj":"EnjuParser_T37","obj":"EnjuParser_T36"},{"id":"EnjuParser_R37","pred":"arg1Of","subj":"EnjuParser_T37","obj":"EnjuParser_T38"},{"id":"EnjuParser_R38","pred":"arg1Of","subj":"EnjuParser_T38","obj":"EnjuParser_T39"},{"id":"EnjuParser_R39","pred":"arg2Of","subj":"EnjuParser_T41","obj":"EnjuParser_T39"},{"id":"EnjuParser_R40","pred":"arg1Of","subj":"EnjuParser_T41","obj":"EnjuParser_T40"},{"id":"EnjuParser_R41","pred":"arg2Of","subj":"EnjuParser_T41","obj":"EnjuParser_T42"},{"id":"EnjuParser_R42","pred":"arg3Of","subj":"EnjuParser_T47","obj":"EnjuParser_T42"},{"id":"EnjuParser_R43","pred":"arg1Of","subj":"EnjuParser_T43","obj":"EnjuParser_T44"},{"id":"EnjuParser_R44","pred":"arg2Of","subj":"EnjuParser_T45","obj":"EnjuParser_T44"},{"id":"EnjuParser_R45","pred":"arg1Of","subj":"EnjuParser_T47","obj":"EnjuParser_T46"},{"id":"EnjuParser_R46","pred":"arg1Of","subj":"EnjuParser_T44","obj":"EnjuParser_T47"},{"id":"EnjuParser_R47","pred":"arg2Of","subj":"EnjuParser_T48","obj":"EnjuParser_T47"},{"id":"EnjuParser_R48","pred":"arg1Of","subj":"EnjuParser_T51","obj":"EnjuParser_T49"},{"id":"EnjuParser_R49","pred":"arg1Of","subj":"EnjuParser_T51","obj":"EnjuParser_T50"},{"id":"EnjuParser_R50","pred":"arg1Of","subj":"EnjuParser_T51","obj":"EnjuParser_T52"},{"id":"EnjuParser_R51","pred":"arg2Of","subj":"EnjuParser_T54","obj":"EnjuParser_T52"},{"id":"EnjuParser_R52","pred":"arg1Of","subj":"EnjuParser_T54","obj":"EnjuParser_T53"},{"id":"EnjuParser_R53","pred":"arg1Of","subj":"EnjuParser_T51","obj":"EnjuParser_T55"},{"id":"EnjuParser_R54","pred":"arg2Of","subj":"EnjuParser_T56","obj":"EnjuParser_T55"},{"id":"EnjuParser_R55","pred":"arg1Of","subj":"EnjuParser_T75","obj":"EnjuParser_T56"},{"id":"EnjuParser_R56","pred":"arg2Of","subj":"EnjuParser_T51","obj":"EnjuParser_T56"},{"id":"EnjuParser_R57","pred":"arg1Of","subj":"EnjuParser_T56","obj":"EnjuParser_T57"},{"id":"EnjuParser_R58","pred":"arg2Of","subj":"EnjuParser_T59","obj":"EnjuParser_T57"},{"id":"EnjuParser_R59","pred":"arg1Of","subj":"EnjuParser_T59","obj":"EnjuParser_T58"},{"id":"EnjuParser_R60","pred":"arg1Of","subj":"EnjuParser_T56","obj":"EnjuParser_T60"},{"id":"EnjuParser_R61","pred":"arg1Of","subj":"EnjuParser_T56","obj":"EnjuParser_T61"},{"id":"EnjuParser_R62","pred":"arg2Of","subj":"EnjuParser_T62","obj":"EnjuParser_T61"},{"id":"EnjuParser_R63","pred":"arg1Of","subj":"EnjuParser_T62","obj":"EnjuParser_T63"},{"id":"EnjuParser_R64","pred":"arg2Of","subj":"EnjuParser_T64","obj":"EnjuParser_T63"},{"id":"EnjuParser_R65","pred":"arg2Of","subj":"EnjuParser_T75","obj":"EnjuParser_T65"},{"id":"EnjuParser_R66","pred":"arg1Of","subj":"EnjuParser_T66","obj":"EnjuParser_T67"},{"id":"EnjuParser_R67","pred":"arg2Of","subj":"EnjuParser_T69","obj":"EnjuParser_T67"},{"id":"EnjuParser_R68","pred":"arg1Of","subj":"EnjuParser_T69","obj":"EnjuParser_T68"},{"id":"EnjuParser_R69","pred":"arg1Of","subj":"EnjuParser_T69","obj":"EnjuParser_T70"},{"id":"EnjuParser_R70","pred":"arg2Of","subj":"EnjuParser_T71","obj":"EnjuParser_T70"},{"id":"EnjuParser_R71","pred":"arg1Of","subj":"EnjuParser_T71","obj":"EnjuParser_T72"},{"id":"EnjuParser_R72","pred":"arg2Of","subj":"EnjuParser_T73","obj":"EnjuParser_T72"},{"id":"EnjuParser_R73","pred":"arg1Of","subj":"EnjuParser_T75","obj":"EnjuParser_T74"},{"id":"EnjuParser_R74","pred":"arg1Of","subj":"EnjuParser_T66","obj":"EnjuParser_T75"},{"id":"EnjuParser_R75","pred":"arg2Of","subj":"EnjuParser_T76","obj":"EnjuParser_T75"},{"id":"EnjuParser_R76","pred":"arg1Of","subj":"EnjuParser_T76","obj":"EnjuParser_T77"},{"id":"EnjuParser_R77","pred":"arg2Of","subj":"EnjuParser_T78","obj":"EnjuParser_T77"},{"id":"EnjuParser_R78","pred":"arg1Of","subj":"EnjuParser_T78","obj":"EnjuParser_T79"},{"id":"EnjuParser_R79","pred":"arg2Of","subj":"EnjuParser_T80","obj":"EnjuParser_T79"},{"id":"EnjuParser_R80","pred":"arg3Of","subj":"EnjuParser_T81","obj":"EnjuParser_T79"},{"id":"EnjuParser_R81","pred":"arg1Of","subj":"EnjuParser_T83","obj":"EnjuParser_T82"},{"id":"EnjuParser_R82","pred":"arg1Of","subj":"EnjuParser_T83","obj":"EnjuParser_T84"},{"id":"EnjuParser_R83","pred":"arg2Of","subj":"EnjuParser_T86","obj":"EnjuParser_T84"},{"id":"EnjuParser_R84","pred":"arg1Of","subj":"EnjuParser_T86","obj":"EnjuParser_T85"},{"id":"EnjuParser_R85","pred":"arg1Of","subj":"EnjuParser_T83","obj":"EnjuParser_T87"},{"id":"EnjuParser_R86","pred":"arg2Of","subj":"EnjuParser_T89","obj":"EnjuParser_T87"},{"id":"EnjuParser_R87","pred":"arg1Of","subj":"EnjuParser_T83","obj":"EnjuParser_T88"},{"id":"EnjuParser_R88","pred":"arg2Of","subj":"EnjuParser_T89","obj":"EnjuParser_T88"},{"id":"EnjuParser_R89","pred":"arg2Of","subj":"EnjuParser_T83","obj":"EnjuParser_T89"},{"id":"EnjuParser_R90","pred":"arg1Of","subj":"EnjuParser_T89","obj":"EnjuParser_T90"},{"id":"EnjuParser_R91","pred":"arg2Of","subj":"EnjuParser_T105","obj":"EnjuParser_T90"},{"id":"EnjuParser_R92","pred":"arg1Of","subj":"EnjuParser_T93","obj":"EnjuParser_T92"},{"id":"EnjuParser_R93","pred":"arg1Of","subj":"EnjuParser_T91","obj":"EnjuParser_T93"},{"id":"EnjuParser_R94","pred":"arg2Of","subj":"EnjuParser_T100","obj":"EnjuParser_T93"},{"id":"EnjuParser_R95","pred":"arg1Of","subj":"EnjuParser_T94","obj":"EnjuParser_T95"},{"id":"EnjuParser_R96","pred":"arg2Of","subj":"EnjuParser_T96","obj":"EnjuParser_T95"},{"id":"EnjuParser_R97","pred":"arg1Of","subj":"EnjuParser_T95","obj":"EnjuParser_T97"},{"id":"EnjuParser_R98","pred":"arg2Of","subj":"EnjuParser_T98","obj":"EnjuParser_T97"},{"id":"EnjuParser_R99","pred":"arg1Of","subj":"EnjuParser_T100","obj":"EnjuParser_T99"},{"id":"EnjuParser_R100","pred":"arg1Of","subj":"EnjuParser_T97","obj":"EnjuParser_T100"},{"id":"EnjuParser_R101","pred":"arg2Of","subj":"EnjuParser_T103","obj":"EnjuParser_T100"},{"id":"EnjuParser_R102","pred":"arg1Of","subj":"EnjuParser_T103","obj":"EnjuParser_T101"},{"id":"EnjuParser_R103","pred":"arg1Of","subj":"EnjuParser_T103","obj":"EnjuParser_T102"},{"id":"EnjuParser_R104","pred":"arg1Of","subj":"EnjuParser_T91","obj":"EnjuParser_T104"},{"id":"EnjuParser_R105","pred":"arg2Of","subj":"EnjuParser_T105","obj":"EnjuParser_T104"},{"id":"EnjuParser_R106","pred":"arg2Of","subj":"EnjuParser_T91","obj":"EnjuParser_T105"},{"id":"EnjuParser_R107","pred":"arg1Of","subj":"EnjuParser_T105","obj":"EnjuParser_T106"},{"id":"EnjuParser_R108","pred":"arg2Of","subj":"EnjuParser_T108","obj":"EnjuParser_T106"},{"id":"EnjuParser_R109","pred":"arg1Of","subj":"EnjuParser_T108","obj":"EnjuParser_T107"},{"id":"EnjuParser_R110","pred":"arg1Of","subj":"EnjuParser_T121","obj":"EnjuParser_T109"},{"id":"EnjuParser_R111","pred":"arg1Of","subj":"EnjuParser_T121","obj":"EnjuParser_T110"},{"id":"EnjuParser_R112","pred":"arg1Of","subj":"EnjuParser_T112","obj":"EnjuParser_T111"},{"id":"EnjuParser_R113","pred":"arg1Of","subj":"EnjuParser_T112","obj":"EnjuParser_T113"},{"id":"EnjuParser_R114","pred":"arg2Of","subj":"EnjuParser_T116","obj":"EnjuParser_T113"},{"id":"EnjuParser_R115","pred":"arg1Of","subj":"EnjuParser_T116","obj":"EnjuParser_T114"},{"id":"EnjuParser_R116","pred":"arg1Of","subj":"EnjuParser_T116","obj":"EnjuParser_T115"},{"id":"EnjuParser_R117","pred":"arg1Of","subj":"EnjuParser_T112","obj":"EnjuParser_T117"},{"id":"EnjuParser_R118","pred":"arg2Of","subj":"EnjuParser_T120","obj":"EnjuParser_T117"},{"id":"EnjuParser_R119","pred":"arg1Of","subj":"EnjuParser_T120","obj":"EnjuParser_T118"},{"id":"EnjuParser_R120","pred":"arg1Of","subj":"EnjuParser_T120","obj":"EnjuParser_T119"},{"id":"EnjuParser_R121","pred":"arg1Of","subj":"EnjuParser_T112","obj":"EnjuParser_T121"},{"id":"EnjuParser_R122","pred":"arg1Of","subj":"EnjuParser_T121","obj":"EnjuParser_T122"},{"id":"EnjuParser_R123","pred":"arg2Of","subj":"EnjuParser_T123","obj":"EnjuParser_T122"},{"id":"EnjuParser_R124","pred":"arg1Of","subj":"EnjuParser_T123","obj":"EnjuParser_T124"},{"id":"EnjuParser_R125","pred":"arg2Of","subj":"EnjuParser_T125","obj":"EnjuParser_T124"},{"id":"EnjuParser_R126","pred":"arg1Of","subj":"EnjuParser_T129","obj":"EnjuParser_T126"},{"id":"EnjuParser_R127","pred":"modOf","subj":"EnjuParser_T133","obj":"EnjuParser_T126"},{"id":"EnjuParser_R128","pred":"arg1Of","subj":"EnjuParser_T126","obj":"EnjuParser_T127"},{"id":"EnjuParser_R129","pred":"arg1Of","subj":"EnjuParser_T128","obj":"EnjuParser_T129"},{"id":"EnjuParser_R130","pred":"arg2Of","subj":"EnjuParser_T130","obj":"EnjuParser_T129"},{"id":"EnjuParser_R131","pred":"arg1Of","subj":"EnjuParser_T129","obj":"EnjuParser_T131"},{"id":"EnjuParser_R132","pred":"arg2Of","subj":"EnjuParser_T132","obj":"EnjuParser_T131"},{"id":"EnjuParser_R133","pred":"arg1Of","subj":"EnjuParser_T129","obj":"EnjuParser_T132"},{"id":"EnjuParser_R134","pred":"arg1Of","subj":"EnjuParser_T131","obj":"EnjuParser_T133"},{"id":"EnjuParser_R135","pred":"arg2Of","subj":"EnjuParser_T136","obj":"EnjuParser_T133"},{"id":"EnjuParser_R136","pred":"arg1Of","subj":"EnjuParser_T129","obj":"EnjuParser_T134"},{"id":"EnjuParser_R137","pred":"arg2Of","subj":"EnjuParser_T136","obj":"EnjuParser_T134"},{"id":"EnjuParser_R138","pred":"arg1Of","subj":"EnjuParser_T129","obj":"EnjuParser_T135"},{"id":"EnjuParser_R139","pred":"arg2Of","subj":"EnjuParser_T136","obj":"EnjuParser_T135"},{"id":"EnjuParser_R140","pred":"arg2Of","subj":"EnjuParser_T129","obj":"EnjuParser_T136"},{"id":"EnjuParser_R141","pred":"arg1Of","subj":"EnjuParser_T138","obj":"EnjuParser_T137"},{"id":"EnjuParser_R142","pred":"arg1Of","subj":"EnjuParser_T136","obj":"EnjuParser_T138"},{"id":"EnjuParser_R143","pred":"arg2Of","subj":"EnjuParser_T140","obj":"EnjuParser_T138"},{"id":"EnjuParser_R144","pred":"arg1Of","subj":"EnjuParser_T140","obj":"EnjuParser_T139"},{"id":"EnjuParser_R145","pred":"arg1Of","subj":"EnjuParser_T162","obj":"EnjuParser_T141"},{"id":"EnjuParser_R146","pred":"arg1Of","subj":"EnjuParser_T162","obj":"EnjuParser_T142"},{"id":"EnjuParser_R147","pred":"arg1Of","subj":"EnjuParser_T144","obj":"EnjuParser_T143"},{"id":"EnjuParser_R148","pred":"arg1Of","subj":"EnjuParser_T144","obj":"EnjuParser_T145"},{"id":"EnjuParser_R149","pred":"arg2Of","subj":"EnjuParser_T146","obj":"EnjuParser_T145"},{"id":"EnjuParser_R150","pred":"arg1Of","subj":"EnjuParser_T148","obj":"EnjuParser_T147"},{"id":"EnjuParser_R151","pred":"arg1Of","subj":"EnjuParser_T146","obj":"EnjuParser_T148"},{"id":"EnjuParser_R152","pred":"arg2Of","subj":"EnjuParser_T155","obj":"EnjuParser_T148"},{"id":"EnjuParser_R153","pred":"arg1Of","subj":"EnjuParser_T150","obj":"EnjuParser_T149"},{"id":"EnjuParser_R154","pred":"arg1Of","subj":"EnjuParser_T150","obj":"EnjuParser_T151"},{"id":"EnjuParser_R155","pred":"arg2Of","subj":"EnjuParser_T153","obj":"EnjuParser_T151"},{"id":"EnjuParser_R156","pred":"arg1Of","subj":"EnjuParser_T153","obj":"EnjuParser_T152"},{"id":"EnjuParser_R157","pred":"arg1Of","subj":"EnjuParser_T155","obj":"EnjuParser_T154"},{"id":"EnjuParser_R158","pred":"arg1Of","subj":"EnjuParser_T151","obj":"EnjuParser_T155"},{"id":"EnjuParser_R159","pred":"arg2Of","subj":"EnjuParser_T157","obj":"EnjuParser_T155"},{"id":"EnjuParser_R160","pred":"arg1Of","subj":"EnjuParser_T157","obj":"EnjuParser_T156"},{"id":"EnjuParser_R161","pred":"arg1Of","subj":"EnjuParser_T144","obj":"EnjuParser_T158"},{"id":"EnjuParser_R162","pred":"arg2Of","subj":"EnjuParser_T159","obj":"EnjuParser_T158"},{"id":"EnjuParser_R163","pred":"arg2Of","subj":"EnjuParser_T160","obj":"EnjuParser_T159"},{"id":"EnjuParser_R164","pred":"arg1Of","subj":"EnjuParser_T162","obj":"EnjuParser_T161"},{"id":"EnjuParser_R165","pred":"arg1Of","subj":"EnjuParser_T144","obj":"EnjuParser_T162"},{"id":"EnjuParser_R166","pred":"arg2Of","subj":"EnjuParser_T165","obj":"EnjuParser_T162"},{"id":"EnjuParser_R167","pred":"arg1Of","subj":"EnjuParser_T165","obj":"EnjuParser_T163"},{"id":"EnjuParser_R168","pred":"arg1Of","subj":"EnjuParser_T165","obj":"EnjuParser_T164"},{"id":"EnjuParser_R169","pred":"arg1Of","subj":"EnjuParser_T165","obj":"EnjuParser_T166"},{"id":"EnjuParser_R170","pred":"arg2Of","subj":"EnjuParser_T167","obj":"EnjuParser_T166"},{"id":"EnjuParser_R171","pred":"arg1Of","subj":"EnjuParser_T169","obj":"EnjuParser_T168"},{"id":"EnjuParser_R172","pred":"arg2Of","subj":"EnjuParser_T169","obj":"EnjuParser_T170"},{"id":"EnjuParser_R173","pred":"arg1Of","subj":"EnjuParser_T169","obj":"EnjuParser_T171"},{"id":"EnjuParser_R174","pred":"arg2Of","subj":"EnjuParser_T173","obj":"EnjuParser_T171"},{"id":"EnjuParser_R175","pred":"arg1Of","subj":"EnjuParser_T173","obj":"EnjuParser_T172"},{"id":"EnjuParser_R176","pred":"arg1Of","subj":"EnjuParser_T169","obj":"EnjuParser_T173"},{"id":"EnjuParser_R177","pred":"arg2Of","subj":"EnjuParser_T175","obj":"EnjuParser_T173"},{"id":"EnjuParser_R178","pred":"arg1Of","subj":"EnjuParser_T175","obj":"EnjuParser_T174"},{"id":"EnjuParser_R179","pred":"arg1Of","subj":"EnjuParser_T177","obj":"EnjuParser_T176"},{"id":"EnjuParser_R180","pred":"modOf","subj":"EnjuParser_T175","obj":"EnjuParser_T176"},{"id":"EnjuParser_R181","pred":"arg2Of","subj":"EnjuParser_T180","obj":"EnjuParser_T177"},{"id":"EnjuParser_R182","pred":"arg1Of","subj":"EnjuParser_T180","obj":"EnjuParser_T178"},{"id":"EnjuParser_R183","pred":"arg1Of","subj":"EnjuParser_T180","obj":"EnjuParser_T179"},{"id":"EnjuParser_R184","pred":"arg1Of","subj":"EnjuParser_T180","obj":"EnjuParser_T181"},{"id":"EnjuParser_R185","pred":"arg2Of","subj":"EnjuParser_T183","obj":"EnjuParser_T181"},{"id":"EnjuParser_R186","pred":"arg1Of","subj":"EnjuParser_T183","obj":"EnjuParser_T182"},{"id":"EnjuParser_R187","pred":"arg1Of","subj":"EnjuParser_T204","obj":"EnjuParser_T184"},{"id":"EnjuParser_R188","pred":"arg2Of","subj":"EnjuParser_T186","obj":"EnjuParser_T184"},{"id":"EnjuParser_R189","pred":"arg1Of","subj":"EnjuParser_T186","obj":"EnjuParser_T185"},{"id":"EnjuParser_R190","pred":"arg2Of","subj":"EnjuParser_T186","obj":"EnjuParser_T187"},{"id":"EnjuParser_R191","pred":"arg1Of","subj":"EnjuParser_T204","obj":"EnjuParser_T188"},{"id":"EnjuParser_R192","pred":"arg1Of","subj":"EnjuParser_T189","obj":"EnjuParser_T190"},{"id":"EnjuParser_R193","pred":"arg1Of","subj":"EnjuParser_T189","obj":"EnjuParser_T191"},{"id":"EnjuParser_R194","pred":"arg2Of","subj":"EnjuParser_T193","obj":"EnjuParser_T191"},{"id":"EnjuParser_R195","pred":"arg1Of","subj":"EnjuParser_T193","obj":"EnjuParser_T192"},{"id":"EnjuParser_R196","pred":"arg1Of","subj":"EnjuParser_T191","obj":"EnjuParser_T194"},{"id":"EnjuParser_R197","pred":"arg2Of","subj":"EnjuParser_T196","obj":"EnjuParser_T194"},{"id":"EnjuParser_R198","pred":"arg1Of","subj":"EnjuParser_T196","obj":"EnjuParser_T195"},{"id":"EnjuParser_R199","pred":"arg1Of","subj":"EnjuParser_T194","obj":"EnjuParser_T197"},{"id":"EnjuParser_R200","pred":"arg2Of","subj":"EnjuParser_T199","obj":"EnjuParser_T197"},{"id":"EnjuParser_R201","pred":"arg1Of","subj":"EnjuParser_T199","obj":"EnjuParser_T198"},{"id":"EnjuParser_R202","pred":"arg1Of","subj":"EnjuParser_T201","obj":"EnjuParser_T200"},{"id":"EnjuParser_R203","pred":"arg1Of","subj":"EnjuParser_T197","obj":"EnjuParser_T201"},{"id":"EnjuParser_R204","pred":"arg2Of","subj":"EnjuParser_T203","obj":"EnjuParser_T201"},{"id":"EnjuParser_R205","pred":"arg1Of","subj":"EnjuParser_T203","obj":"EnjuParser_T202"},{"id":"EnjuParser_R206","pred":"arg1Of","subj":"EnjuParser_T201","obj":"EnjuParser_T204"},{"id":"EnjuParser_R207","pred":"arg2Of","subj":"EnjuParser_T206","obj":"EnjuParser_T204"},{"id":"EnjuParser_R208","pred":"arg1Of","subj":"EnjuParser_T206","obj":"EnjuParser_T205"},{"id":"EnjuParser_R209","pred":"arg1Of","subj":"EnjuParser_T201","obj":"EnjuParser_T206"},{"id":"EnjuParser_R210","pred":"arg1Of","subj":"EnjuParser_T206","obj":"EnjuParser_T207"},{"id":"EnjuParser_R211","pred":"arg2Of","subj":"EnjuParser_T209","obj":"EnjuParser_T207"},{"id":"EnjuParser_R212","pred":"arg1Of","subj":"EnjuParser_T209","obj":"EnjuParser_T208"},{"id":"EnjuParser_R213","pred":"arg1Of","subj":"EnjuParser_T209","obj":"EnjuParser_T210"},{"id":"EnjuParser_R214","pred":"arg2Of","subj":"EnjuParser_T217","obj":"EnjuParser_T210"},{"id":"EnjuParser_R215","pred":"arg1Of","subj":"EnjuParser_T215","obj":"EnjuParser_T211"},{"id":"EnjuParser_R216","pred":"arg1Of","subj":"EnjuParser_T215","obj":"EnjuParser_T212"},{"id":"EnjuParser_R217","pred":"arg1Of","subj":"EnjuParser_T212","obj":"EnjuParser_T213"},{"id":"EnjuParser_R218","pred":"arg2Of","subj":"EnjuParser_T214","obj":"EnjuParser_T213"},{"id":"EnjuParser_R219","pred":"arg1Of","subj":"EnjuParser_T215","obj":"EnjuParser_T214"},{"id":"EnjuParser_R220","pred":"arg1Of","subj":"EnjuParser_T217","obj":"EnjuParser_T216"},{"id":"EnjuParser_R221","pred":"arg1Of","subj":"EnjuParser_T215","obj":"EnjuParser_T217"},{"id":"EnjuParser_R222","pred":"arg2Of","subj":"EnjuParser_T218","obj":"EnjuParser_T217"},{"id":"EnjuParser_R223","pred":"arg1Of","subj":"EnjuParser_T218","obj":"EnjuParser_T219"},{"id":"EnjuParser_R224","pred":"arg1Of","subj":"EnjuParser_T204","obj":"EnjuParser_T220"},{"id":"EnjuParser_R225","pred":"arg1Of","subj":"EnjuParser_T204","obj":"EnjuParser_T221"},{"id":"EnjuParser_R226","pred":"arg2Of","subj":"EnjuParser_T228","obj":"EnjuParser_T221"},{"id":"EnjuParser_R227","pred":"arg1Of","subj":"EnjuParser_T222","obj":"EnjuParser_T223"},{"id":"EnjuParser_R228","pred":"arg2Of","subj":"EnjuParser_T224","obj":"EnjuParser_T223"},{"id":"EnjuParser_R229","pred":"arg1Of","subj":"EnjuParser_T226","obj":"EnjuParser_T225"},{"id":"EnjuParser_R230","pred":"arg1Of","subj":"EnjuParser_T223","obj":"EnjuParser_T226"},{"id":"EnjuParser_R231","pred":"arg2Of","subj":"EnjuParser_T227","obj":"EnjuParser_T226"},{"id":"EnjuParser_R232","pred":"arg1Of","subj":"EnjuParser_T226","obj":"EnjuParser_T228"},{"id":"EnjuParser_R233","pred":"arg2Of","subj":"EnjuParser_T230","obj":"EnjuParser_T228"},{"id":"EnjuParser_R234","pred":"arg1Of","subj":"EnjuParser_T230","obj":"EnjuParser_T229"},{"id":"EnjuParser_R235","pred":"arg1Of","subj":"EnjuParser_T232","obj":"EnjuParser_T231"},{"id":"EnjuParser_R236","pred":"arg1Of","subj":"EnjuParser_T232","obj":"EnjuParser_T233"},{"id":"EnjuParser_R237","pred":"arg1Of","subj":"EnjuParser_T233","obj":"EnjuParser_T234"},{"id":"EnjuParser_R238","pred":"arg2Of","subj":"EnjuParser_T236","obj":"EnjuParser_T234"},{"id":"EnjuParser_R239","pred":"arg1Of","subj":"EnjuParser_T235","obj":"EnjuParser_T236"},{"id":"EnjuParser_R240","pred":"arg2Of","subj":"EnjuParser_T237","obj":"EnjuParser_T236"},{"id":"EnjuParser_R241","pred":"arg1Of","subj":"EnjuParser_T232","obj":"EnjuParser_T238"},{"id":"EnjuParser_R242","pred":"arg2Of","subj":"EnjuParser_T240","obj":"EnjuParser_T238"},{"id":"EnjuParser_R243","pred":"arg1Of","subj":"EnjuParser_T232","obj":"EnjuParser_T239"},{"id":"EnjuParser_R244","pred":"arg2Of","subj":"EnjuParser_T240","obj":"EnjuParser_T239"},{"id":"EnjuParser_R245","pred":"arg2Of","subj":"EnjuParser_T232","obj":"EnjuParser_T240"},{"id":"EnjuParser_R246","pred":"arg1Of","subj":"EnjuParser_T240","obj":"EnjuParser_T241"},{"id":"EnjuParser_R247","pred":"arg2Of","subj":"EnjuParser_T250","obj":"EnjuParser_T241"},{"id":"EnjuParser_R248","pred":"arg1Of","subj":"EnjuParser_T241","obj":"EnjuParser_T242"},{"id":"EnjuParser_R249","pred":"arg1Of","subj":"EnjuParser_T248","obj":"EnjuParser_T243"},{"id":"EnjuParser_R250","pred":"arg1Of","subj":"EnjuParser_T248","obj":"EnjuParser_T244"},{"id":"EnjuParser_R251","pred":"arg1Of","subj":"EnjuParser_T248","obj":"EnjuParser_T245"},{"id":"EnjuParser_R252","pred":"arg1Of","subj":"EnjuParser_T245","obj":"EnjuParser_T246"},{"id":"EnjuParser_R253","pred":"arg2Of","subj":"EnjuParser_T247","obj":"EnjuParser_T246"},{"id":"EnjuParser_R254","pred":"arg1Of","subj":"EnjuParser_T248","obj":"EnjuParser_T247"},{"id":"EnjuParser_R255","pred":"arg1Of","subj":"EnjuParser_T248","obj":"EnjuParser_T249"},{"id":"EnjuParser_R256","pred":"arg2Of","subj":"EnjuParser_T250","obj":"EnjuParser_T249"},{"id":"EnjuParser_R257","pred":"arg2Of","subj":"EnjuParser_T248","obj":"EnjuParser_T250"},{"id":"EnjuParser_R258","pred":"arg1Of","subj":"EnjuParser_T251","obj":"EnjuParser_T252"},{"id":"EnjuParser_R259","pred":"arg2Of","subj":"EnjuParser_T253","obj":"EnjuParser_T252"},{"id":"EnjuParser_R260","pred":"arg1Of","subj":"EnjuParser_T251","obj":"EnjuParser_T254"},{"id":"EnjuParser_R261","pred":"arg2Of","subj":"EnjuParser_T255","obj":"EnjuParser_T254"},{"id":"EnjuParser_R262","pred":"arg1Of","subj":"EnjuParser_T251","obj":"EnjuParser_T256"},{"id":"EnjuParser_R263","pred":"arg2Of","subj":"EnjuParser_T259","obj":"EnjuParser_T256"},{"id":"EnjuParser_R264","pred":"arg1Of","subj":"EnjuParser_T259","obj":"EnjuParser_T257"},{"id":"EnjuParser_R265","pred":"arg1Of","subj":"EnjuParser_T259","obj":"EnjuParser_T258"},{"id":"EnjuParser_R266","pred":"arg1Of","subj":"EnjuParser_T259","obj":"EnjuParser_T260"},{"id":"EnjuParser_R267","pred":"arg2Of","subj":"EnjuParser_T262","obj":"EnjuParser_T260"},{"id":"EnjuParser_R268","pred":"arg1Of","subj":"EnjuParser_T262","obj":"EnjuParser_T261"},{"id":"EnjuParser_R269","pred":"arg1Of","subj":"EnjuParser_T266","obj":"EnjuParser_T263"},{"id":"EnjuParser_R270","pred":"arg1Of","subj":"EnjuParser_T266","obj":"EnjuParser_T264"},{"id":"EnjuParser_R271","pred":"arg1Of","subj":"EnjuParser_T266","obj":"EnjuParser_T265"},{"id":"EnjuParser_R272","pred":"arg1Of","subj":"EnjuParser_T266","obj":"EnjuParser_T267"},{"id":"EnjuParser_R273","pred":"arg2Of","subj":"EnjuParser_T268","obj":"EnjuParser_T267"},{"id":"EnjuParser_R274","pred":"arg3Of","subj":"EnjuParser_T269","obj":"EnjuParser_T267"},{"id":"EnjuParser_R275","pred":"arg1Of","subj":"EnjuParser_T266","obj":"EnjuParser_T270"},{"id":"EnjuParser_R276","pred":"arg2Of","subj":"EnjuParser_T271","obj":"EnjuParser_T270"},{"id":"EnjuParser_R277","pred":"arg1Of","subj":"EnjuParser_T266","obj":"EnjuParser_T272"},{"id":"EnjuParser_R278","pred":"arg2Of","subj":"EnjuParser_T266","obj":"EnjuParser_T273"},{"id":"EnjuParser_R279","pred":"arg1Of","subj":"EnjuParser_T273","obj":"EnjuParser_T274"},{"id":"EnjuParser_R280","pred":"arg2Of","subj":"EnjuParser_T276","obj":"EnjuParser_T274"},{"id":"EnjuParser_R281","pred":"arg1Of","subj":"EnjuParser_T276","obj":"EnjuParser_T275"},{"id":"EnjuParser_R282","pred":"arg1Of","subj":"EnjuParser_T276","obj":"EnjuParser_T277"},{"id":"EnjuParser_R283","pred":"arg2Of","subj":"EnjuParser_T278","obj":"EnjuParser_T277"},{"id":"EnjuParser_R284","pred":"arg2Of","subj":"EnjuParser_T276","obj":"EnjuParser_T279"},{"id":"EnjuParser_R285","pred":"arg1Of","subj":"EnjuParser_T279","obj":"EnjuParser_T280"},{"id":"EnjuParser_R286","pred":"arg2Of","subj":"EnjuParser_T281","obj":"EnjuParser_T280"},{"id":"EnjuParser_R287","pred":"arg1Of","subj":"EnjuParser_T283","obj":"EnjuParser_T282"},{"id":"EnjuParser_R288","pred":"arg1Of","subj":"EnjuParser_T266","obj":"EnjuParser_T283"},{"id":"EnjuParser_R289","pred":"arg2Of","subj":"EnjuParser_T285","obj":"EnjuParser_T283"},{"id":"EnjuParser_R290","pred":"arg1Of","subj":"EnjuParser_T285","obj":"EnjuParser_T284"},{"id":"EnjuParser_R291","pred":"arg1Of","subj":"EnjuParser_T283","obj":"EnjuParser_T286"},{"id":"EnjuParser_R292","pred":"arg2Of","subj":"EnjuParser_T287","obj":"EnjuParser_T286"},{"id":"EnjuParser_R293","pred":"arg1Of","subj":"EnjuParser_T290","obj":"EnjuParser_T287"},{"id":"EnjuParser_R294","pred":"arg2Of","subj":"EnjuParser_T290","obj":"EnjuParser_T288"},{"id":"EnjuParser_R295","pred":"arg1Of","subj":"EnjuParser_T290","obj":"EnjuParser_T289"},{"id":"EnjuParser_R296","pred":"arg1Of","subj":"EnjuParser_T290","obj":"EnjuParser_T291"},{"id":"EnjuParser_R297","pred":"arg2Of","subj":"EnjuParser_T292","obj":"EnjuParser_T291"},{"id":"EnjuParser_R298","pred":"arg1Of","subj":"EnjuParser_T291","obj":"EnjuParser_T293"},{"id":"EnjuParser_R299","pred":"arg2Of","subj":"EnjuParser_T294","obj":"EnjuParser_T293"},{"id":"EnjuParser_R300","pred":"arg1Of","subj":"EnjuParser_T290","obj":"EnjuParser_T294"},{"id":"EnjuParser_R301","pred":"arg2Of","subj":"EnjuParser_T296","obj":"EnjuParser_T294"},{"id":"EnjuParser_R302","pred":"arg1Of","subj":"EnjuParser_T296","obj":"EnjuParser_T295"},{"id":"EnjuParser_R303","pred":"arg1Of","subj":"EnjuParser_T298","obj":"EnjuParser_T297"},{"id":"EnjuParser_R304","pred":"arg1Of","subj":"EnjuParser_T298","obj":"EnjuParser_T299"},{"id":"EnjuParser_R305","pred":"arg2Of","subj":"EnjuParser_T301","obj":"EnjuParser_T299"},{"id":"EnjuParser_R306","pred":"arg2Of","subj":"EnjuParser_T301","obj":"EnjuParser_T300"},{"id":"EnjuParser_R307","pred":"arg1Of","subj":"EnjuParser_T298","obj":"EnjuParser_T302"},{"id":"EnjuParser_R308","pred":"arg2Of","subj":"EnjuParser_T303","obj":"EnjuParser_T302"},{"id":"EnjuParser_R309","pred":"arg1Of","subj":"EnjuParser_T298","obj":"EnjuParser_T304"},{"id":"EnjuParser_R310","pred":"arg2Of","subj":"EnjuParser_T306","obj":"EnjuParser_T304"},{"id":"EnjuParser_R311","pred":"arg1Of","subj":"EnjuParser_T304","obj":"EnjuParser_T305"},{"id":"EnjuParser_R312","pred":"arg1Of","subj":"EnjuParser_T298","obj":"EnjuParser_T306"},{"id":"EnjuParser_R313","pred":"arg1Of","subj":"EnjuParser_T306","obj":"EnjuParser_T307"},{"id":"EnjuParser_R314","pred":"arg2Of","subj":"EnjuParser_T309","obj":"EnjuParser_T307"},{"id":"EnjuParser_R315","pred":"arg1Of","subj":"EnjuParser_T309","obj":"EnjuParser_T308"},{"id":"EnjuParser_R316","pred":"arg1Of","subj":"EnjuParser_T309","obj":"EnjuParser_T310"},{"id":"EnjuParser_R317","pred":"arg2Of","subj":"EnjuParser_T312","obj":"EnjuParser_T310"},{"id":"EnjuParser_R318","pred":"arg1Of","subj":"EnjuParser_T312","obj":"EnjuParser_T311"},{"id":"EnjuParser_R319","pred":"arg1Of","subj":"EnjuParser_T313","obj":"EnjuParser_T314"},{"id":"EnjuParser_R320","pred":"arg2Of","subj":"EnjuParser_T328","obj":"EnjuParser_T314"},{"id":"EnjuParser_R321","pred":"arg1Of","subj":"EnjuParser_T328","obj":"EnjuParser_T315"},{"id":"EnjuParser_R322","pred":"arg1Of","subj":"EnjuParser_T316","obj":"EnjuParser_T317"},{"id":"EnjuParser_R323","pred":"arg2Of","subj":"EnjuParser_T319","obj":"EnjuParser_T317"},{"id":"EnjuParser_R324","pred":"arg1Of","subj":"EnjuParser_T319","obj":"EnjuParser_T318"},{"id":"EnjuParser_R325","pred":"arg1Of","subj":"EnjuParser_T319","obj":"EnjuParser_T320"},{"id":"EnjuParser_R326","pred":"arg2Of","subj":"EnjuParser_T321","obj":"EnjuParser_T320"},{"id":"EnjuParser_R327","pred":"arg1Of","subj":"EnjuParser_T316","obj":"EnjuParser_T322"},{"id":"EnjuParser_R328","pred":"arg2Of","subj":"EnjuParser_T323","obj":"EnjuParser_T322"},{"id":"EnjuParser_R329","pred":"arg1Of","subj":"EnjuParser_T323","obj":"EnjuParser_T324"},{"id":"EnjuParser_R330","pred":"arg2Of","subj":"EnjuParser_T327","obj":"EnjuParser_T324"},{"id":"EnjuParser_R331","pred":"arg1Of","subj":"EnjuParser_T327","obj":"EnjuParser_T325"},{"id":"EnjuParser_R332","pred":"arg1Of","subj":"EnjuParser_T327","obj":"EnjuParser_T326"},{"id":"EnjuParser_R333","pred":"arg1Of","subj":"EnjuParser_T316","obj":"EnjuParser_T328"},{"id":"EnjuParser_R334","pred":"arg2Of","subj":"EnjuParser_T330","obj":"EnjuParser_T328"},{"id":"EnjuParser_R335","pred":"arg1Of","subj":"EnjuParser_T328","obj":"EnjuParser_T329"},{"id":"EnjuParser_R336","pred":"arg1Of","subj":"EnjuParser_T316","obj":"EnjuParser_T330"},{"id":"EnjuParser_R337","pred":"arg1Of","subj":"EnjuParser_T330","obj":"EnjuParser_T331"},{"id":"EnjuParser_R338","pred":"arg2Of","subj":"EnjuParser_T333","obj":"EnjuParser_T331"},{"id":"EnjuParser_R339","pred":"arg1Of","subj":"EnjuParser_T333","obj":"EnjuParser_T332"},{"id":"EnjuParser_R340","pred":"arg1Of","subj":"EnjuParser_T333","obj":"EnjuParser_T334"},{"id":"EnjuParser_R341","pred":"arg2Of","subj":"EnjuParser_T342","obj":"EnjuParser_T334"},{"id":"EnjuParser_R342","pred":"arg1Of","subj":"EnjuParser_T342","obj":"EnjuParser_T335"},{"id":"EnjuParser_R343","pred":"arg1Of","subj":"EnjuParser_T342","obj":"EnjuParser_T336"},{"id":"EnjuParser_R344","pred":"arg1Of","subj":"EnjuParser_T342","obj":"EnjuParser_T337"},{"id":"EnjuParser_R345","pred":"arg1Of","subj":"EnjuParser_T342","obj":"EnjuParser_T338"},{"id":"EnjuParser_R346","pred":"arg1Of","subj":"EnjuParser_T340","obj":"EnjuParser_T339"},{"id":"EnjuParser_R347","pred":"arg1Of","subj":"EnjuParser_T338","obj":"EnjuParser_T340"},{"id":"EnjuParser_R348","pred":"arg2Of","subj":"EnjuParser_T341","obj":"EnjuParser_T340"},{"id":"EnjuParser_R349","pred":"arg1Of","subj":"EnjuParser_T342","obj":"EnjuParser_T341"},{"id":"EnjuParser_R350","pred":"arg1Of","subj":"EnjuParser_T342","obj":"EnjuParser_T343"},{"id":"EnjuParser_R351","pred":"arg2Of","subj":"EnjuParser_T345","obj":"EnjuParser_T343"},{"id":"EnjuParser_R352","pred":"arg1Of","subj":"EnjuParser_T345","obj":"EnjuParser_T344"}],"namespaces":[{"prefix":"_base","uri":"http://kmcs.nii.ac.jp/enju/"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
performance-test
{"project":"performance-test","denotations":[{"id":"PD-UBERON-AE-B_T1","span":{"begin":734,"end":738},"obj":"http://purl.obolibrary.org/obo/UBERON_0001913"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
NCBITAXON
{"project":"NCBITAXON","denotations":[{"id":"T1","span":{"begin":61,"end":64},"obj":"OrganismTaxon"}],"attributes":[{"id":"A1","pred":"db_id","subj":"T1","obj":"10114"},{"id":"A2","pred":"db_id","subj":"T1","obj":"10116"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}
Anatomy-UBERON
{"project":"Anatomy-UBERON","denotations":[{"id":"T1","span":{"begin":721,"end":724},"obj":"Body_part"},{"id":"T2","span":{"begin":734,"end":738},"obj":"Body_part"},{"id":"T3","span":{"begin":2162,"end":2173},"obj":"Body_part"},{"id":"T4","span":{"begin":2175,"end":2188},"obj":"Body_part"}],"attributes":[{"id":"A1","pred":"uberon_id","subj":"T1","obj":"http://purl.obolibrary.org/obo/UBERON_0001013"},{"id":"A2","pred":"uberon_id","subj":"T2","obj":"http://purl.obolibrary.org/obo/UBERON_0001913"},{"id":"A3","pred":"uberon_id","subj":"T3","obj":"http://purl.obolibrary.org/obo/GO_0005737"},{"id":"A4","pred":"uberon_id","subj":"T4","obj":"http://purl.obolibrary.org/obo/GO_0016020"}],"text":"Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis.\nRat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity-purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand-binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit."}