VEGF165-binding sites within heparan sulfate encompass two highly sulfated domains and can be liberated by K5 lyase.

2.50
Hdl Handle:
http://hdl.handle.net/10541/72698
Title:
VEGF165-binding sites within heparan sulfate encompass two highly sulfated domains and can be liberated by K5 lyase.
Authors:
Robinson, Christopher J; Mulloy, Barbara; Gallagher, John T; Stringer, Sally E
Abstract:
The vascular endothelial growth factor (VEGF) family of proteins controls the formation and growth of blood vessels. The most potent and widely expressed isoform, VEGF165, is secreted as a disulfide-linked homodimer with two identical heparin-binding sites. Interactions with heparan sulfate (HS) regulate the diffusion, half-life, and affinity of VEGF165 for its signaling receptors. We have determined a number of key HS structural features that mediate the specific binding of the VEGF165 dimer. Carboxylate groups and 2-O-, 6-O-, and N-sulfation of HS contributed to the strength of the VEGF165 interaction; however, 6-O-sulfates appeared to be particularly important. Cleavage of HS by heparinase, heparitinase, or heparanase severely reduced VEGF165 binding. In contrast, K5 lyase-cleaved HS retained significant VEGF165 affinity, suggesting that binding sites for the growth factor are present within extended stretches of sulfation. Binding studies and molecular modeling demonstrated that an oligosaccharide 6 or 7 residues long was sufficient to fully occupy the heparin-binding site of a VEGF165 monomer. The data presented are consistent with a model whereby the two heparin-binding sites of the VEGF165 dimer interact simultaneously with highly sulfated S-domain regions of the HS chain that can be linked through a stretch of transition sequence.
Affiliation:
Cancer Research UK and University of Manchester Department of Medical Oncology, Christie Hospital National Health Service Trust, Wilmslow Road, Manchester M20 4BX, United Kindgom. Christopher.Robinson@manchester.ac.uk
Citation:
VEGF165-binding sites within heparan sulfate encompass two highly sulfated domains and can be liberated by K5 lyase. 2006, 281 (3):1731-40 J. Biol. Chem.
Journal:
The Journal of Biological Chemistry
Issue Date:
20-Jan-2006
URI:
http://hdl.handle.net/10541/72698
DOI:
10.1074/jbc.M510760200
PubMed ID:
16258170
Type:
Article
Language:
en
ISSN:
0021-9258
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorRobinson, Christopher J-
dc.contributor.authorMulloy, Barbara-
dc.contributor.authorGallagher, John T-
dc.contributor.authorStringer, Sally E-
dc.date.accessioned2009-07-07T09:55:47Z-
dc.date.available2009-07-07T09:55:47Z-
dc.date.issued2006-01-20-
dc.identifier.citationVEGF165-binding sites within heparan sulfate encompass two highly sulfated domains and can be liberated by K5 lyase. 2006, 281 (3):1731-40 J. Biol. Chem.en
dc.identifier.issn0021-9258-
dc.identifier.pmid16258170-
dc.identifier.doi10.1074/jbc.M510760200-
dc.identifier.urihttp://hdl.handle.net/10541/72698-
dc.description.abstractThe vascular endothelial growth factor (VEGF) family of proteins controls the formation and growth of blood vessels. The most potent and widely expressed isoform, VEGF165, is secreted as a disulfide-linked homodimer with two identical heparin-binding sites. Interactions with heparan sulfate (HS) regulate the diffusion, half-life, and affinity of VEGF165 for its signaling receptors. We have determined a number of key HS structural features that mediate the specific binding of the VEGF165 dimer. Carboxylate groups and 2-O-, 6-O-, and N-sulfation of HS contributed to the strength of the VEGF165 interaction; however, 6-O-sulfates appeared to be particularly important. Cleavage of HS by heparinase, heparitinase, or heparanase severely reduced VEGF165 binding. In contrast, K5 lyase-cleaved HS retained significant VEGF165 affinity, suggesting that binding sites for the growth factor are present within extended stretches of sulfation. Binding studies and molecular modeling demonstrated that an oligosaccharide 6 or 7 residues long was sufficient to fully occupy the heparin-binding site of a VEGF165 monomer. The data presented are consistent with a model whereby the two heparin-binding sites of the VEGF165 dimer interact simultaneously with highly sulfated S-domain regions of the HS chain that can be linked through a stretch of transition sequence.en
dc.language.isoenen
dc.subject.mesh3T3 Cells-
dc.subject.meshAnimals-
dc.subject.meshCarbohydrate Conformation-
dc.subject.meshCattle-
dc.subject.meshChromatography, High Pressure Liquid-
dc.subject.meshDimerization-
dc.subject.meshDisaccharides-
dc.subject.meshGlycosaminoglycans-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshMice-
dc.subject.meshModels, Molecular-
dc.subject.meshRecombinant Proteins-
dc.subject.meshSharks-
dc.subject.meshVascular Endothelial Growth Factor A-
dc.titleVEGF165-binding sites within heparan sulfate encompass two highly sulfated domains and can be liberated by K5 lyase.en
dc.typeArticleen
dc.contributor.departmentCancer Research UK and University of Manchester Department of Medical Oncology, Christie Hospital National Health Service Trust, Wilmslow Road, Manchester M20 4BX, United Kindgom. Christopher.Robinson@manchester.ac.uken
dc.identifier.journalThe Journal of Biological Chemistryen

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