A new model for the domain structure of heparan sulfate based on the novel specificity of K5 lyase.

2.50
Hdl Handle:
http://hdl.handle.net/10541/78452
Title:
A new model for the domain structure of heparan sulfate based on the novel specificity of K5 lyase.
Authors:
Murphy, Kevin J; Merry, Catherine L R; Lyon, Malcolm; Thompson, James E; Roberts, Ian S; Gallagher, John T
Abstract:
Elucidation of the molecular structure of heparan sulfate (HS) is the key to understanding its functional versatility as a co-receptor for growth factors and morphogens. We have identified and exploited the novel substrate specificity of the coliphage K5 lyase in studies of the domain organization of HS. We show that K5 lyase cleaves HS principally within non-sulfated sequences of four or more N-acetylated disaccharides. Uniquely, sections comprising alternating N-acetylated and N-sulfated units are resistant to the enzyme, as are the highly sulfated S domains. Spacing of the K5 lyase cleavage sites ( approximately 7-8 kDa) is similar to that of the S domains. On the basis of these findings, we propose a refined model of the structure of HS in which N-acetylated sequences of four to five disaccharide units (GlcNAc-GlcUA)(4-5) are positioned centrally between the S domains. The latter are embedded within N-acetylated and N-sulfated sequences, forming extended regions of hypervariable sulfation distributed at regular intervals along the polymer chain. K5 lyase provides a means of excision of these composite sulfated regions for structural and functional analyses.
Affiliation:
Cancer Research UK and University of Manchester, Department of Medical Oncology, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, United Kingdom.
Citation:
A new model for the domain structure of heparan sulfate based on the novel specificity of K5 lyase. 2004, 279 (26):27239-45 J. Biol. Chem.
Journal:
The Journal of Biological Chemistry
Issue Date:
25-Jun-2004
URI:
http://hdl.handle.net/10541/78452
DOI:
10.1074/jbc.M401774200
PubMed ID:
15047699
Type:
Article
Language:
en
ISSN:
0021-9258
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorMurphy, Kevin J-
dc.contributor.authorMerry, Catherine L R-
dc.contributor.authorLyon, Malcolm-
dc.contributor.authorThompson, James E-
dc.contributor.authorRoberts, Ian S-
dc.contributor.authorGallagher, John T-
dc.date.accessioned2009-08-25T11:25:49Z-
dc.date.available2009-08-25T11:25:49Z-
dc.date.issued2004-06-25-
dc.identifier.citationA new model for the domain structure of heparan sulfate based on the novel specificity of K5 lyase. 2004, 279 (26):27239-45 J. Biol. Chem.en
dc.identifier.issn0021-9258-
dc.identifier.pmid15047699-
dc.identifier.doi10.1074/jbc.M401774200-
dc.identifier.urihttp://hdl.handle.net/10541/78452-
dc.description.abstractElucidation of the molecular structure of heparan sulfate (HS) is the key to understanding its functional versatility as a co-receptor for growth factors and morphogens. We have identified and exploited the novel substrate specificity of the coliphage K5 lyase in studies of the domain organization of HS. We show that K5 lyase cleaves HS principally within non-sulfated sequences of four or more N-acetylated disaccharides. Uniquely, sections comprising alternating N-acetylated and N-sulfated units are resistant to the enzyme, as are the highly sulfated S domains. Spacing of the K5 lyase cleavage sites ( approximately 7-8 kDa) is similar to that of the S domains. On the basis of these findings, we propose a refined model of the structure of HS in which N-acetylated sequences of four to five disaccharide units (GlcNAc-GlcUA)(4-5) are positioned centrally between the S domains. The latter are embedded within N-acetylated and N-sulfated sequences, forming extended regions of hypervariable sulfation distributed at regular intervals along the polymer chain. K5 lyase provides a means of excision of these composite sulfated regions for structural and functional analyses.en
dc.language.isoenen
dc.subject.meshChromatography, Gel-
dc.subject.meshDisaccharides-
dc.subject.meshEscherichia coli Proteins-
dc.subject.meshFlavobacterium-
dc.subject.meshGlucosamine-
dc.subject.meshHeparin Lyase-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshModels, Molecular-
dc.subject.meshNitrous Acid-
dc.subject.meshOligosaccharides-
dc.subject.meshPolysaccharide-Lyases-
dc.subject.meshProtein Structure, Tertiary-
dc.subject.meshRecombinant Proteins-
dc.subject.meshSubstrate Specificity-
dc.subject.meshTritium-
dc.titleA new model for the domain structure of heparan sulfate based on the novel specificity of K5 lyase.en
dc.typeArticleen
dc.contributor.departmentCancer Research UK and University of Manchester, Department of Medical Oncology, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, United Kingdom.en
dc.identifier.journalThe Journal of Biological Chemistryen
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