Neuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain.

2.50
Hdl Handle:
http://hdl.handle.net/10541/80056
Title:
Neuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain.
Authors:
Mamluk, Roni; Gechtman, Ze'ev; Kutcher, Matthew E; Gasiunas, Nijole; Gallagher, John T; Klagsbrun, Michael
Abstract:
Neuroplin-1 (NRP1), a receptor for vascular endothelial growth factor (VEGF) family members, has three distinct extracellular domains, a1a2, b1b2, and c. To determine the VEGF(165) and placenta growth factor 2 (PlGF-2)-binding sites of NRP1, recombinant NRP1 domains were expressed in mammalian cells as Myc-tagged, soluble proteins, and used in co-precipitation experiments with 125I-VEGF165 and 125I-PlGF-2. Anti-Myc antibodies immunoprecipitated 125I-VEGF165 and 125I-PlGF-2 in the presence of the b1b2 but not of the a1a2 and c domains. Neither b1 nor b2 alone was capable of binding 125I-VEGF165. In competition experiments, VEGF165 competed PlGF-2 binding to the NRP1 b1b2 domain, suggesting that the binding sites of VEGF165 and PlGF-2 overlap. The presence of the a1a2 domain greatly enhanced VEGF165, but not PlGF-2 binding to b1b2. Heparin enhanced the binding of both 125I-VEGF165 and 125I-PlGF-2 to the b1b2 domain by 20- and 4-fold, respectively. A heparin chain of at least 20-24 monosaccharides was necessary for binding. In addition, the b1b2 domain of NRP1 could bind heparin directly, requiring heparin oligomers of at least 8 monosaccharide units. It was concluded that an intact b1b2 domain serves as the VEGF165-, PlGF-2-, and heparin-binding sites in NRP1, and that heparin is a critical component for regulating VEGF165 and PlGF-2 interactions with NRP1 by physically interacting with both receptor and ligands.
Affiliation:
Department of Surgical Research, Children's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.
Citation:
Neuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain. 2002, 277 (27):24818-25 J. Biol. Chem.
Journal:
The Journal of Biological Chemistry
Issue Date:
5-Jul-2002
URI:
http://hdl.handle.net/10541/80056
DOI:
10.1074/jbc.M200730200
PubMed ID:
11986311
Type:
Article
Language:
en
ISSN:
0021-9258
Appears in Collections:
All Christie Publications

Full metadata record

DC FieldValue Language
dc.contributor.authorMamluk, Roni-
dc.contributor.authorGechtman, Ze'ev-
dc.contributor.authorKutcher, Matthew E-
dc.contributor.authorGasiunas, Nijole-
dc.contributor.authorGallagher, John T-
dc.contributor.authorKlagsbrun, Michael-
dc.date.accessioned2009-09-07T12:49:28Z-
dc.date.available2009-09-07T12:49:28Z-
dc.date.issued2002-07-05-
dc.identifier.citationNeuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain. 2002, 277 (27):24818-25 J. Biol. Chem.en
dc.identifier.issn0021-9258-
dc.identifier.pmid11986311-
dc.identifier.doi10.1074/jbc.M200730200-
dc.identifier.urihttp://hdl.handle.net/10541/80056-
dc.description.abstractNeuroplin-1 (NRP1), a receptor for vascular endothelial growth factor (VEGF) family members, has three distinct extracellular domains, a1a2, b1b2, and c. To determine the VEGF(165) and placenta growth factor 2 (PlGF-2)-binding sites of NRP1, recombinant NRP1 domains were expressed in mammalian cells as Myc-tagged, soluble proteins, and used in co-precipitation experiments with 125I-VEGF165 and 125I-PlGF-2. Anti-Myc antibodies immunoprecipitated 125I-VEGF165 and 125I-PlGF-2 in the presence of the b1b2 but not of the a1a2 and c domains. Neither b1 nor b2 alone was capable of binding 125I-VEGF165. In competition experiments, VEGF165 competed PlGF-2 binding to the NRP1 b1b2 domain, suggesting that the binding sites of VEGF165 and PlGF-2 overlap. The presence of the a1a2 domain greatly enhanced VEGF165, but not PlGF-2 binding to b1b2. Heparin enhanced the binding of both 125I-VEGF165 and 125I-PlGF-2 to the b1b2 domain by 20- and 4-fold, respectively. A heparin chain of at least 20-24 monosaccharides was necessary for binding. In addition, the b1b2 domain of NRP1 could bind heparin directly, requiring heparin oligomers of at least 8 monosaccharide units. It was concluded that an intact b1b2 domain serves as the VEGF165-, PlGF-2-, and heparin-binding sites in NRP1, and that heparin is a critical component for regulating VEGF165 and PlGF-2 interactions with NRP1 by physically interacting with both receptor and ligands.en
dc.language.isoenen
dc.subjectBreast Canceren
dc.subjectCultured Tumour Cellsen
dc.subject.meshAngiogenesis Inducing Agents-
dc.subject.meshBase Sequence-
dc.subject.meshBinding Sites-
dc.subject.meshBreast Neoplasms-
dc.subject.meshCells, Cultured-
dc.subject.meshCloning, Molecular-
dc.subject.meshDNA Primers-
dc.subject.meshEndothelial Growth Factors-
dc.subject.meshEndothelium, Vascular-
dc.subject.meshHeparin-
dc.subject.meshHumans-
dc.subject.meshKinetics-
dc.subject.meshLymphokines-
dc.subject.meshNerve Tissue Proteins-
dc.subject.meshNeuropilin-1-
dc.subject.meshPregnancy Proteins-
dc.subject.meshRecombinant Proteins-
dc.subject.meshTumor Cells, Cultured-
dc.subject.meshVascular Endothelial Growth Factor A-
dc.subject.meshVascular Endothelial Growth Factors-
dc.titleNeuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain.en
dc.typeArticleen
dc.contributor.departmentDepartment of Surgical Research, Children's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.en
dc.identifier.journalThe Journal of Biological Chemistryen

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