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PMID:10471394

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Citation

Herley, MT, Yu, Y, Whitney, RG and Sato, JD (1999) Characterization of the VEGF binding site on the Flt-1 receptor. Biochem. Biophys. Res. Commun. 262:731-8

Abstract

The angiogenic growth factor VEGF binds to the receptor tyrosine kinases Flt-1 and KDR/Flk-1. Immunoglobulin (Ig)-like loop-2 of Flt-1 is involved in binding VEGF, but the contribution of other Flt-1 Ig-loops to VEGF binding remains unclear. We tested the ability of membrane-bound chimeras between the extracellular domain of Flt-1 and the cell adhesion molecule embigin to bind VEGF. VEGF bound as well to receptors containing Flt-1 loops 1-2 or 2-3 as it did to the entire Flt-1 extracellular domain. Chimeras containing only loop-2 of Flt-1 bound VEGF with 22-fold lower affinity. We conclude that high-affinity VEGF binding requires Ig-like loop-2 plus either loop-1 or loop-3. In addition, Flt-1 amino acid residues Arg-224 and Asp-231 were not essential for high-affinity binding of VEGF to membrane-bound Flt-1.

Links

PubMed Online version:10.1006/bbrc.1999.1282

Keywords

Amino Acid Substitution; Animals; Base Sequence; Binding Sites; Cloning, Molecular; DNA Primers; Endothelial Growth Factors/metabolism; Glycoproteins/chemistry; Glycoproteins/metabolism; Humans; Kinetics; Lymphokines/metabolism; Molecular Sequence Data; Mutagenesis, Site-Directed; Protein Structure, Secondary; Proto-Oncogene Proteins/chemistry; Proto-Oncogene Proteins/genetics; Proto-Oncogene Proteins/metabolism; Rats; Receptor Protein-Tyrosine Kinases/chemistry; Receptor Protein-Tyrosine Kinases/genetics; Receptor Protein-Tyrosine Kinases/metabolism; Receptors, Growth Factor/chemistry; Receptors, Growth Factor/genetics; Receptors, Growth Factor/metabolism; Receptors, Vascular Endothelial Growth Factor; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/metabolism; Recombinant Proteins/metabolism; Restriction Mapping; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor Receptor-1; Vascular Endothelial Growth Factors

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