Distinct substrate specificities of bacterial heparinases against N-unsubstituted glucosamine residues in heparan sulfate.

2.50
Hdl Handle:
http://hdl.handle.net/10541/75661
Title:
Distinct substrate specificities of bacterial heparinases against N-unsubstituted glucosamine residues in heparan sulfate.
Authors:
Wei, Zheng; Lyon, Malcolm; Gallagher, John T
Abstract:
The rare N-unsubstituted glucosamine (GlcNH(3)(+)) residues in heparan sulfate have important biological and pathophysiological roles. In this study, four GlcNH(3)(+)-containing disaccharides were obtained from partially de-N-sulfated forms of heparin and the N-sulfated K5 polysaccharide by digestion with combined heparinases I, II, and III. These were identified as DeltaHexA-GlcNH(3)(+),DeltaHexA-GlcNH(3)(+)(6S),DeltaHexA(2S)-GlcNH(3)(+), and DeltaHexA(2S)-GlcNH(3)(+)(6S). Digestions with individual enzymes revealed that heparinase I did not cleave at GlcNH(3)(+) residues; however, heparinases II and III showed selective and distinct activities. Heparinase II generated DeltaHexA-GlcNH(3)(+)(6S),DeltaHexA(2S)-GlcNH(3)(+), and DeltaHexA(2S)-GlcNH(3)(+)(6S) disaccharides, whereas heparinase III yielded only the DeltaHexA-GlcNH(3)(+) unit. Thus, the action of heparinase II requires O-sulfation, whereas heparinase III acts only on the corresponding non-sulfated unit. These striking distinctions in substrate specificities of heparinases could be used to isolate oligosaccharides with novel sequences of GlcNH(3)(+) residues. Finally, heparinases were used to identify and quantify GlcNH(3)(+)-containing disaccharides in native bovine kidney and porcine intestinal mucosal heparan sulfates. The relatively high content of O-sulfated GlcNH(3)(+)-disaccharides in kidney HS raises questions about how these sequences are generated.
Affiliation:
Cancer Research UK and the University of Manchester Department of Medical Oncology, Christie Hospital National Health Service Trust, Wilmslow Road, Manchester M20 4BX, United Kingdom.
Citation:
Distinct substrate specificities of bacterial heparinases against N-unsubstituted glucosamine residues in heparan sulfate. 2005, 280 (16):15742-8 J. Biol. Chem.
Journal:
The Journal of Biological Chemistry
Issue Date:
22-Apr-2005
URI:
http://hdl.handle.net/10541/75661
DOI:
10.1074/jbc.M501102200
PubMed ID:
15705564
Type:
Article
Language:
en
ISSN:
0021-9258
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorWei, Zheng-
dc.contributor.authorLyon, Malcolm-
dc.contributor.authorGallagher, John T-
dc.date.accessioned2009-07-24T15:36:00Z-
dc.date.available2009-07-24T15:36:00Z-
dc.date.issued2005-04-22-
dc.identifier.citationDistinct substrate specificities of bacterial heparinases against N-unsubstituted glucosamine residues in heparan sulfate. 2005, 280 (16):15742-8 J. Biol. Chem.en
dc.identifier.issn0021-9258-
dc.identifier.pmid15705564-
dc.identifier.doi10.1074/jbc.M501102200-
dc.identifier.urihttp://hdl.handle.net/10541/75661-
dc.description.abstractThe rare N-unsubstituted glucosamine (GlcNH(3)(+)) residues in heparan sulfate have important biological and pathophysiological roles. In this study, four GlcNH(3)(+)-containing disaccharides were obtained from partially de-N-sulfated forms of heparin and the N-sulfated K5 polysaccharide by digestion with combined heparinases I, II, and III. These were identified as DeltaHexA-GlcNH(3)(+),DeltaHexA-GlcNH(3)(+)(6S),DeltaHexA(2S)-GlcNH(3)(+), and DeltaHexA(2S)-GlcNH(3)(+)(6S). Digestions with individual enzymes revealed that heparinase I did not cleave at GlcNH(3)(+) residues; however, heparinases II and III showed selective and distinct activities. Heparinase II generated DeltaHexA-GlcNH(3)(+)(6S),DeltaHexA(2S)-GlcNH(3)(+), and DeltaHexA(2S)-GlcNH(3)(+)(6S) disaccharides, whereas heparinase III yielded only the DeltaHexA-GlcNH(3)(+) unit. Thus, the action of heparinase II requires O-sulfation, whereas heparinase III acts only on the corresponding non-sulfated unit. These striking distinctions in substrate specificities of heparinases could be used to isolate oligosaccharides with novel sequences of GlcNH(3)(+) residues. Finally, heparinases were used to identify and quantify GlcNH(3)(+)-containing disaccharides in native bovine kidney and porcine intestinal mucosal heparan sulfates. The relatively high content of O-sulfated GlcNH(3)(+)-disaccharides in kidney HS raises questions about how these sequences are generated.en
dc.language.isoenen
dc.subject.meshChromatography, High Pressure Liquid-
dc.subject.meshFlavobacterium-
dc.subject.meshGlucosamine-
dc.subject.meshHeparin Lyase-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshPolysaccharide-Lyases-
dc.subject.meshSubstrate Specificity-
dc.subject.meshTime Factors-
dc.titleDistinct substrate specificities of bacterial heparinases against N-unsubstituted glucosamine residues in heparan sulfate.en
dc.typeArticleen
dc.contributor.departmentCancer Research UK and the University of Manchester Department of Medical Oncology, Christie Hospital National Health Service Trust, Wilmslow Road, Manchester M20 4BX, United Kingdom.en
dc.identifier.journalThe Journal of Biological Chemistryen

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