PubMed:10988250 JSONTXT

{"project":"Glycan-Motif","denotations":[{"id":"T1","span":{"begin":74,"end":84},"obj":"https://glytoucan.org/Structures/Glycans/G00017MO"},{"id":"T2","span":{"begin":198,"end":208},"obj":"https://glytoucan.org/Structures/Glycans/G00017MO"},{"id":"T3","span":{"begin":289,"end":299},"obj":"https://glytoucan.org/Structures/Glycans/G00017MO"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"GlyCosmos6-Glycan-Motif-Image","denotations":[{"id":"T1","span":{"begin":74,"end":84},"obj":"Glycan_Motif"},{"id":"T2","span":{"begin":198,"end":208},"obj":"Glycan_Motif"},{"id":"T3","span":{"begin":289,"end":299},"obj":"Glycan_Motif"}],"attributes":[{"id":"A1","pred":"image","subj":"T1","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G00017MO"},{"id":"A2","pred":"image","subj":"T2","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G00017MO"},{"id":"A3","pred":"image","subj":"T3","obj":"https://api.glycosmos.org/wurcs2image/0.10.0/png/binary/G00017MO"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"sentences","denotations":[{"id":"TextSentencer_T1","span":{"begin":0,"end":137},"obj":"Sentence"},{"id":"TextSentencer_T2","span":{"begin":138,"end":245},"obj":"Sentence"},{"id":"TextSentencer_T3","span":{"begin":246,"end":381},"obj":"Sentence"},{"id":"TextSentencer_T4","span":{"begin":382,"end":467},"obj":"Sentence"},{"id":"TextSentencer_T5","span":{"begin":468,"end":607},"obj":"Sentence"},{"id":"TextSentencer_T6","span":{"begin":608,"end":689},"obj":"Sentence"},{"id":"TextSentencer_T7","span":{"begin":690,"end":840},"obj":"Sentence"},{"id":"TextSentencer_T8","span":{"begin":841,"end":965},"obj":"Sentence"},{"id":"TextSentencer_T9","span":{"begin":966,"end":1119},"obj":"Sentence"},{"id":"TextSentencer_T10","span":{"begin":1120,"end":1310},"obj":"Sentence"},{"id":"TextSentencer_T11","span":{"begin":1311,"end":1452},"obj":"Sentence"},{"id":"TextSentencer_T12","span":{"begin":1453,"end":1579},"obj":"Sentence"},{"id":"T1","span":{"begin":0,"end":137},"obj":"Sentence"},{"id":"T2","span":{"begin":138,"end":245},"obj":"Sentence"},{"id":"T3","span":{"begin":246,"end":381},"obj":"Sentence"},{"id":"T4","span":{"begin":382,"end":467},"obj":"Sentence"},{"id":"T5","span":{"begin":468,"end":607},"obj":"Sentence"},{"id":"T6","span":{"begin":608,"end":689},"obj":"Sentence"},{"id":"T7","span":{"begin":690,"end":840},"obj":"Sentence"},{"id":"T8","span":{"begin":841,"end":965},"obj":"Sentence"},{"id":"T9","span":{"begin":966,"end":1119},"obj":"Sentence"},{"id":"T10","span":{"begin":1120,"end":1310},"obj":"Sentence"},{"id":"T11","span":{"begin":1311,"end":1452},"obj":"Sentence"},{"id":"T12","span":{"begin":1453,"end":1579},"obj":"Sentence"},{"id":"T1","span":{"begin":0,"end":137},"obj":"Sentence"},{"id":"T2","span":{"begin":138,"end":245},"obj":"Sentence"},{"id":"T3","span":{"begin":246,"end":381},"obj":"Sentence"},{"id":"T4","span":{"begin":382,"end":467},"obj":"Sentence"},{"id":"T5","span":{"begin":468,"end":607},"obj":"Sentence"},{"id":"T6","span":{"begin":608,"end":689},"obj":"Sentence"},{"id":"T7","span":{"begin":690,"end":840},"obj":"Sentence"},{"id":"T8","span":{"begin":841,"end":965},"obj":"Sentence"},{"id":"T9","span":{"begin":966,"end":1119},"obj":"Sentence"},{"id":"T10","span":{"begin":1120,"end":1310},"obj":"Sentence"},{"id":"T11","span":{"begin":1311,"end":1452},"obj":"Sentence"},{"id":"T12","span":{"begin":1453,"end":1579},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"GlyCosmos6-Glycan-Motif-Structure","denotations":[{"id":"T1","span":{"begin":74,"end":84},"obj":"https://glytoucan.org/Structures/Glycans/G00017MO"},{"id":"T2","span":{"begin":198,"end":208},"obj":"https://glytoucan.org/Structures/Glycans/G00017MO"},{"id":"T3","span":{"begin":289,"end":299},"obj":"https://glytoucan.org/Structures/Glycans/G00017MO"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"PubmedHPO","denotations":[{"id":"T1","span":{"begin":728,"end":738},"obj":"HP_0009609"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"GlycoBiology-FMA","denotations":[{"id":"_T1","span":{"begin":74,"end":84},"obj":"FMAID:63014"},{"id":"_T2","span":{"begin":74,"end":84},"obj":"FMAID:167396"},{"id":"_T3","span":{"begin":198,"end":208},"obj":"FMAID:167396"},{"id":"_T4","span":{"begin":198,"end":208},"obj":"FMAID:63014"},{"id":"_T5","span":{"begin":214,"end":221},"obj":"FMAID:199625"},{"id":"_T6","span":{"begin":214,"end":221},"obj":"FMAID:85272"},{"id":"_T7","span":{"begin":289,"end":299},"obj":"FMAID:167396"},{"id":"_T8","span":{"begin":289,"end":299},"obj":"FMAID:63014"},{"id":"_T9","span":{"begin":885,"end":895},"obj":"FMAID:82750"},{"id":"_T10","span":{"begin":885,"end":895},"obj":"FMAID:196739"},{"id":"_T11","span":{"begin":911,"end":917},"obj":"FMAID:82758"},{"id":"_T12","span":{"begin":911,"end":917},"obj":"FMAID:196747"},{"id":"_T13","span":{"begin":1354,"end":1362},"obj":"FMAID:67257"},{"id":"_T14","span":{"begin":1354,"end":1362},"obj":"FMAID:165447"}],"namespaces":[{"prefix":"FMAID","uri":"http://purl.org/sig/ont/fma/fma"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"uniprot-human","denotations":[{"id":"T1","span":{"begin":22,"end":33},"obj":"http://www.uniprot.org/uniprot/Q99484"},{"id":"T2","span":{"begin":668,"end":679},"obj":"http://www.uniprot.org/uniprot/Q99484"},{"id":"T3","span":{"begin":1021,"end":1032},"obj":"http://www.uniprot.org/uniprot/Q99484"},{"id":"T4","span":{"begin":1098,"end":1109},"obj":"http://www.uniprot.org/uniprot/Q99484"},{"id":"T5","span":{"begin":1162,"end":1173},"obj":"http://www.uniprot.org/uniprot/Q99484"},{"id":"T6","span":{"begin":210,"end":212},"obj":"http://www.uniprot.org/uniprot/P69208"},{"id":"T7","span":{"begin":370,"end":372},"obj":"http://www.uniprot.org/uniprot/P69208"},{"id":"T8","span":{"begin":500,"end":502},"obj":"http://www.uniprot.org/uniprot/P69208"},{"id":"T9","span":{"begin":636,"end":638},"obj":"http://www.uniprot.org/uniprot/P69208"},{"id":"T10","span":{"begin":930,"end":932},"obj":"http://www.uniprot.org/uniprot/P69208"},{"id":"T11","span":{"begin":1392,"end":1394},"obj":"http://www.uniprot.org/uniprot/P69208"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"uniprot-mouse","denotations":[{"id":"T1","span":{"begin":22,"end":33},"obj":"http://www.uniprot.org/uniprot/P38649"},{"id":"T2","span":{"begin":668,"end":679},"obj":"http://www.uniprot.org/uniprot/P38649"},{"id":"T3","span":{"begin":1021,"end":1032},"obj":"http://www.uniprot.org/uniprot/P38649"},{"id":"T4","span":{"begin":1098,"end":1109},"obj":"http://www.uniprot.org/uniprot/P38649"},{"id":"T5","span":{"begin":1162,"end":1173},"obj":"http://www.uniprot.org/uniprot/P38649"},{"id":"T6","span":{"begin":772,"end":775},"obj":"http://www.uniprot.org/uniprot/Q5YD48"},{"id":"T7","span":{"begin":772,"end":775},"obj":"http://www.uniprot.org/uniprot/Q9EQ00"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"GlycoBiology-NCBITAXON","denotations":[{"id":"T1","span":{"begin":52,"end":63},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/745"},{"id":"T2","span":{"begin":52,"end":63},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/712"},{"id":"T3","span":{"begin":145,"end":156},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/745"},{"id":"T4","span":{"begin":145,"end":156},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/712"},{"id":"T5","span":{"begin":809,"end":813},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/9973"},{"id":"T6","span":{"begin":814,"end":818},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/158455"},{"id":"T7","span":{"begin":814,"end":818},"obj":"http://purl.bioontology.org/ontology/NCBITAXON/3554"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"GO-BP","denotations":[{"id":"T1","span":{"begin":22,"end":44},"obj":"http://purl.obolibrary.org/obo/GO_0004380"},{"id":"T2","span":{"begin":668,"end":688},"obj":"http://purl.obolibrary.org/obo/GO_0004380"},{"id":"T3","span":{"begin":1021,"end":1041},"obj":"http://purl.obolibrary.org/obo/GO_0004380"},{"id":"T4","span":{"begin":1098,"end":1118},"obj":"http://purl.obolibrary.org/obo/GO_0004380"},{"id":"T5","span":{"begin":1162,"end":1182},"obj":"http://purl.obolibrary.org/obo/GO_0004380"},{"id":"T6","span":{"begin":22,"end":44},"obj":"http://purl.obolibrary.org/obo/GO_0016740"},{"id":"T7","span":{"begin":668,"end":688},"obj":"http://purl.obolibrary.org/obo/GO_0016740"},{"id":"T8","span":{"begin":1021,"end":1041},"obj":"http://purl.obolibrary.org/obo/GO_0016740"},{"id":"T9","span":{"begin":1098,"end":1118},"obj":"http://purl.obolibrary.org/obo/GO_0016740"},{"id":"T10","span":{"begin":1162,"end":1182},"obj":"http://purl.obolibrary.org/obo/GO_0016740"},{"id":"T11","span":{"begin":22,"end":44},"obj":"http://purl.obolibrary.org/obo/GO_0051347"},{"id":"T12","span":{"begin":668,"end":688},"obj":"http://purl.obolibrary.org/obo/GO_0051347"},{"id":"T13","span":{"begin":1021,"end":1041},"obj":"http://purl.obolibrary.org/obo/GO_0051347"},{"id":"T14","span":{"begin":1098,"end":1118},"obj":"http://purl.obolibrary.org/obo/GO_0051347"},{"id":"T15","span":{"begin":1162,"end":1182},"obj":"http://purl.obolibrary.org/obo/GO_0051347"},{"id":"T16","span":{"begin":225,"end":244},"obj":"http://purl.obolibrary.org/obo/GO_0044413"},{"id":"T17","span":{"begin":661,"end":688},"obj":"http://purl.obolibrary.org/obo/GO_0008375"},{"id":"T18","span":{"begin":1091,"end":1118},"obj":"http://purl.obolibrary.org/obo/GO_0008375"},{"id":"T19","span":{"begin":661,"end":688},"obj":"http://purl.obolibrary.org/obo/GO_0016262"},{"id":"T20","span":{"begin":1091,"end":1118},"obj":"http://purl.obolibrary.org/obo/GO_0016262"},{"id":"T21","span":{"begin":933,"end":950},"obj":"http://purl.obolibrary.org/obo/GO_0004517"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"GO-CC","denotations":[{"id":"T1","span":{"begin":231,"end":235},"obj":"http://purl.obolibrary.org/obo/GO_0018995"},{"id":"T2","span":{"begin":269,"end":277},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T3","span":{"begin":410,"end":418},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T4","span":{"begin":565,"end":573},"obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"T5","span":{"begin":780,"end":783},"obj":"http://purl.obolibrary.org/obo/GO_0005790"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"UBERON-AE","denotations":[{"id":"T1","span":{"begin":214,"end":221},"obj":"http://purl.obolibrary.org/obo/UBERON_0003893"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"EDAM-topics","denotations":[{"id":"T1","span":{"begin":178,"end":186},"obj":"http://edamontology.org/topic_0783"},{"id":"T2","span":{"begin":707,"end":715},"obj":"http://edamontology.org/topic_3168"},{"id":"T3","span":{"begin":707,"end":715},"obj":"http://edamontology.org/topic_0080"},{"id":"T4","span":{"begin":765,"end":770},"obj":"http://edamontology.org/topic_0158"},{"id":"T5","span":{"begin":1354,"end":1362},"obj":"http://edamontology.org/topic_0078"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"EDAM-DFO","denotations":[{"id":"T1","span":{"begin":178,"end":186},"obj":"http://edamontology.org/data_3718"},{"id":"T2","span":{"begin":261,"end":268},"obj":"http://edamontology.org/data_1756"},{"id":"T3","span":{"begin":707,"end":715},"obj":"http://edamontology.org/data_2044"},{"id":"T4","span":{"begin":707,"end":715},"obj":"http://edamontology.org/operation_3218"},{"id":"T5","span":{"begin":993,"end":1000},"obj":"http://edamontology.org/data_1756"},{"id":"T6","span":{"begin":1069,"end":1076},"obj":"http://edamontology.org/data_1756"},{"id":"T7","span":{"begin":1209,"end":1216},"obj":"http://edamontology.org/data_3108"},{"id":"T8","span":{"begin":1354,"end":1362},"obj":"http://edamontology.org/data_1467"},{"id":"T9","span":{"begin":1354,"end":1362},"obj":"http://edamontology.org/format_1208"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"NGLY1-deficiency","denotations":[{"id":"PD-NGLY1-deficiency-B_T1","span":{"begin":341,"end":347},"obj":"chem:24139"},{"id":"PD-NGLY1-deficiency-B_T2","span":{"begin":661,"end":667},"obj":"chem:24139"},{"id":"PD-NGLY1-deficiency-B_T3","span":{"begin":1091,"end":1097},"obj":"chem:24139"}],"namespaces":[{"prefix":"hgnc","uri":"https://www.genenames.org/data/gene-symbol-report/#!/hgnc_id/HGNC:"},{"prefix":"omim","uri":"https://www.omim.org/entry/"},{"prefix":"chem","uri":"https://pubchem.ncbi.nlm.nih.gov/compound/"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"GlyTouCan-IUPAC","denotations":[{"id":"GlycanIUPAC_T21","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G89565QL\""},{"id":"GlycanIUPAC_T1","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G26693XF\""},{"id":"GlycanIUPAC_T2","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G26693XF\""},{"id":"GlycanIUPAC_T3","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G26693XF\""},{"id":"GlycanIUPAC_T4","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G01864SU\""},{"id":"GlycanIUPAC_T5","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G01864SU\""},{"id":"GlycanIUPAC_T6","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G01864SU\""},{"id":"GlycanIUPAC_T7","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G17605FD\""},{"id":"GlycanIUPAC_T8","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G17605FD\""},{"id":"GlycanIUPAC_T9","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G17605FD\""},{"id":"GlycanIUPAC_T10","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G41950LU\""},{"id":"GlycanIUPAC_T11","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G41950LU\""},{"id":"GlycanIUPAC_T12","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G41950LU\""},{"id":"GlycanIUPAC_T13","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G57195RJ\""},{"id":"GlycanIUPAC_T14","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G57195RJ\""},{"id":"GlycanIUPAC_T15","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G57195RJ\""},{"id":"GlycanIUPAC_T16","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G85391SA\""},{"id":"GlycanIUPAC_T17","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G85391SA\""},{"id":"GlycanIUPAC_T18","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G85391SA\""},{"id":"GlycanIUPAC_T19","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G89565QL\""},{"id":"GlycanIUPAC_T20","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G89565QL\""},{"id":"GlycanIUPAC_T22","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G80869MR\""},{"id":"GlycanIUPAC_T23","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G80869MR\""},{"id":"GlycanIUPAC_T24","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G80869MR\""},{"id":"GlycanIUPAC_T25","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G55978NL\""},{"id":"GlycanIUPAC_T26","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G55978NL\""},{"id":"GlycanIUPAC_T27","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G55978NL\""},{"id":"GlycanIUPAC_T28","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G54644LT\""},{"id":"GlycanIUPAC_T29","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G54644LT\""},{"id":"GlycanIUPAC_T30","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G54644LT\""},{"id":"GlycanIUPAC_T31","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G25694UG\""},{"id":"GlycanIUPAC_T32","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G25694UG\""},{"id":"GlycanIUPAC_T33","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G25694UG\""},{"id":"GlycanIUPAC_T34","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G25126RB\""},{"id":"GlycanIUPAC_T35","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G25126RB\""},{"id":"GlycanIUPAC_T36","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G25126RB\""},{"id":"GlycanIUPAC_T37","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G51848AD\""},{"id":"GlycanIUPAC_T38","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G51848AD\""},{"id":"GlycanIUPAC_T39","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G51848AD\""},{"id":"GlycanIUPAC_T40","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G94667GM\""},{"id":"GlycanIUPAC_T41","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G94667GM\""},{"id":"GlycanIUPAC_T42","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G94667GM\""},{"id":"GlycanIUPAC_T43","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G30124BO\""},{"id":"GlycanIUPAC_T44","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G30124BO\""},{"id":"GlycanIUPAC_T45","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G30124BO\""},{"id":"GlycanIUPAC_T46","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G82777EZ\""},{"id":"GlycanIUPAC_T47","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G82777EZ\""},{"id":"GlycanIUPAC_T48","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G82777EZ\""},{"id":"GlycanIUPAC_T49","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G10151YZ\""},{"id":"GlycanIUPAC_T50","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G10151YZ\""},{"id":"GlycanIUPAC_T51","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G10151YZ\""},{"id":"GlycanIUPAC_T52","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G17585ZM\""},{"id":"GlycanIUPAC_T53","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G17585ZM\""},{"id":"GlycanIUPAC_T54","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G17585ZM\""},{"id":"GlycanIUPAC_T55","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G04411CJ\""},{"id":"GlycanIUPAC_T56","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G04411CJ\""},{"id":"GlycanIUPAC_T57","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G04411CJ\""},{"id":"GlycanIUPAC_T58","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G38254HJ\""},{"id":"GlycanIUPAC_T59","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G38254HJ\""},{"id":"GlycanIUPAC_T60","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G38254HJ\""},{"id":"GlycanIUPAC_T61","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G75188FS\""},{"id":"GlycanIUPAC_T62","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G75188FS\""},{"id":"GlycanIUPAC_T63","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G75188FS\""},{"id":"GlycanIUPAC_T64","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G70374VG\""},{"id":"GlycanIUPAC_T65","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G70374VG\""},{"id":"GlycanIUPAC_T66","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G70374VG\""},{"id":"GlycanIUPAC_T67","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G45176LJ\""},{"id":"GlycanIUPAC_T68","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G45176LJ\""},{"id":"GlycanIUPAC_T69","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G45176LJ\""},{"id":"GlycanIUPAC_T70","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G30874VW\""},{"id":"GlycanIUPAC_T71","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G30874VW\""},{"id":"GlycanIUPAC_T72","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G30874VW\""},{"id":"GlycanIUPAC_T73","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G69333MI\""},{"id":"GlycanIUPAC_T74","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G69333MI\""},{"id":"GlycanIUPAC_T75","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G69333MI\""},{"id":"GlycanIUPAC_T76","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G10676XO\""},{"id":"GlycanIUPAC_T77","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G10676XO\""},{"id":"GlycanIUPAC_T78","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G10676XO\""},{"id":"GlycanIUPAC_T79","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G14843DJ\""},{"id":"GlycanIUPAC_T80","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G14843DJ\""},{"id":"GlycanIUPAC_T81","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G14843DJ\""},{"id":"GlycanIUPAC_T82","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G47546FR\""},{"id":"GlycanIUPAC_T83","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G47546FR\""},{"id":"GlycanIUPAC_T84","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G47546FR\""},{"id":"GlycanIUPAC_T85","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G73695ZM\""},{"id":"GlycanIUPAC_T86","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G73695ZM\""},{"id":"GlycanIUPAC_T87","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G73695ZM\""},{"id":"GlycanIUPAC_T88","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G31923TJ\""},{"id":"GlycanIUPAC_T89","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G31923TJ\""},{"id":"GlycanIUPAC_T90","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G31923TJ\""},{"id":"GlycanIUPAC_T91","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G60519EP\""},{"id":"GlycanIUPAC_T92","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G60519EP\""},{"id":"GlycanIUPAC_T93","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G60519EP\""},{"id":"GlycanIUPAC_T94","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G07933IA\""},{"id":"GlycanIUPAC_T95","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G07933IA\""},{"id":"GlycanIUPAC_T96","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G07933IA\""},{"id":"GlycanIUPAC_T97","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G40745NH\""},{"id":"GlycanIUPAC_T98","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G40745NH\""},{"id":"GlycanIUPAC_T99","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G40745NH\""},{"id":"GlycanIUPAC_T100","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G54496YV\""},{"id":"GlycanIUPAC_T101","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G54496YV\""},{"id":"GlycanIUPAC_T102","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G54496YV\""},{"id":"GlycanIUPAC_T103","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G62953SQ\""},{"id":"GlycanIUPAC_T104","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G62953SQ\""},{"id":"GlycanIUPAC_T105","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G62953SQ\""},{"id":"GlycanIUPAC_T106","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G70070AY\""},{"id":"GlycanIUPAC_T107","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G70070AY\""},{"id":"GlycanIUPAC_T108","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G70070AY\""},{"id":"GlycanIUPAC_T109","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G78792WC\""},{"id":"GlycanIUPAC_T110","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G78792WC\""},{"id":"GlycanIUPAC_T111","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G78792WC\""},{"id":"GlycanIUPAC_T112","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G25238AV\""},{"id":"GlycanIUPAC_T113","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G25238AV\""},{"id":"GlycanIUPAC_T114","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G25238AV\""},{"id":"GlycanIUPAC_T115","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G40510DP\""},{"id":"GlycanIUPAC_T116","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G40510DP\""},{"id":"GlycanIUPAC_T117","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G40510DP\""},{"id":"GlycanIUPAC_T118","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G61120TK\""},{"id":"GlycanIUPAC_T119","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G61120TK\""},{"id":"GlycanIUPAC_T120","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G61120TK\""},{"id":"GlycanIUPAC_T121","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G41342KV\""},{"id":"GlycanIUPAC_T122","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G41342KV\""},{"id":"GlycanIUPAC_T123","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G41342KV\""},{"id":"GlycanIUPAC_T124","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G90703NA\""},{"id":"GlycanIUPAC_T125","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G90703NA\""},{"id":"GlycanIUPAC_T126","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G90703NA\""},{"id":"GlycanIUPAC_T127","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G01591HR\""},{"id":"GlycanIUPAC_T128","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G01591HR\""},{"id":"GlycanIUPAC_T129","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G01591HR\""},{"id":"GlycanIUPAC_T130","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G56520XN\""},{"id":"GlycanIUPAC_T131","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G56520XN\""},{"id":"GlycanIUPAC_T132","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G56520XN\""},{"id":"GlycanIUPAC_T133","span":{"begin":341,"end":347},"obj":"\"http://rdf.glycoinfo.org/glycan/G81830JX\""},{"id":"GlycanIUPAC_T134","span":{"begin":661,"end":667},"obj":"\"http://rdf.glycoinfo.org/glycan/G81830JX\""},{"id":"GlycanIUPAC_T135","span":{"begin":1091,"end":1097},"obj":"\"http://rdf.glycoinfo.org/glycan/G81830JX\""}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"performance-test","denotations":[{"id":"PD-UBERON-AE-B_T1","span":{"begin":214,"end":221},"obj":"http://purl.obolibrary.org/obo/UBERON_0003893"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"NCBITAXON","denotations":[{"id":"T1","span":{"begin":52,"end":73},"obj":"OrganismTaxon"},{"id":"T2","span":{"begin":145,"end":166},"obj":"OrganismTaxon"}],"attributes":[{"id":"A1","pred":"db_id","subj":"T1","obj":"747"},{"id":"A2","pred":"db_id","subj":"T2","obj":"747"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}

{"project":"Anatomy-UBERON","denotations":[{"id":"T1","span":{"begin":269,"end":277},"obj":"Body_part"},{"id":"T4","span":{"begin":410,"end":418},"obj":"Body_part"},{"id":"T7","span":{"begin":565,"end":573},"obj":"Body_part"}],"attributes":[{"id":"A1","pred":"uberon_id","subj":"T1","obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"A2","pred":"uberon_id","subj":"T1","obj":"http://purl.obolibrary.org/obo/UBERON_0000094"},{"id":"A3","pred":"uberon_id","subj":"T1","obj":"http://purl.obolibrary.org/obo/UBERON_0000158"},{"id":"A4","pred":"uberon_id","subj":"T4","obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"A5","pred":"uberon_id","subj":"T4","obj":"http://purl.obolibrary.org/obo/UBERON_0000094"},{"id":"A6","pred":"uberon_id","subj":"T4","obj":"http://purl.obolibrary.org/obo/UBERON_0000158"},{"id":"A7","pred":"uberon_id","subj":"T7","obj":"http://purl.obolibrary.org/obo/GO_0016020"},{"id":"A8","pred":"uberon_id","subj":"T7","obj":"http://purl.obolibrary.org/obo/UBERON_0000094"},{"id":"A9","pred":"uberon_id","subj":"T7","obj":"http://purl.obolibrary.org/obo/UBERON_0000158"}],"text":"Dissection of the two transferase activities of the Pasteurella multocida hyaluronan synthase: two active sites exist in one polypeptide.\nType A Pasteurella multocida, an animal pathogen, employs a hyaluronan [HA] capsule to avoid host defenses. PmHAS, the 972-residue membrane-associated hyaluronan synthase, catalyzes the transfer of both GlcNAc and GlcUA to form the HA polymer. To define the catalytic and membrane-associated domains, pmHAS mutants were analyzed. PmHAS1-703 is a soluble, active HA synthase suggesting that the carboxyl-terminus is involved in membrane association of the native enzyme. PmHAS1-650 is inactive as a HA synthase, but retains GlcNAc-transferase activity. Within the pmHAS sequence, there is a duplicated domain containing a short motif, Asp-Gly-Ser, that is conserved among many beta-glycosyltransferases. Changing this aspartate in either domain to asparagine, glutamate, or lysine reduced the HA synthase activity to low levels. The mutants substituted at residue 196 possessed GlcUA-transferase activity while those substituted at residue 477 possessed GlcNAc-transferase activity. The Michaelis constants of the functional transferase activity of the various mutants, a measure of the apparent affinity of the enzymes for the precursors, were similar to wild-type values. Furthermore, mixing D196N and D477K mutant proteins in the same reaction allowed HA polymerization at levels similar to the wild-type enzyme. These results provide the first direct evidence that the synthase polypeptide utilizes two separate glycosyltransferase sites."}