PMID:17626017

Citation	Krilleke, D, DeErkenez, A, Schubert, W, Giri, I, Robinson, GS, Ng, YS and Shima, DT (2007) Molecular mapping and functional characterization of the VEGF164 heparin-binding domain. J. Biol. Chem. 282:28045-56
Abstract	The longer splice isoforms of vascular endothelial growth factor-A (VEGF-A), including mouse VEGF164, contain a highly basic heparin-binding domain (HBD), which imparts the ability of these isoforms to be deposited in the heparan sulfate-rich extracellular matrix and to interact with the prototype sulfated glycosaminoglycan, heparin. The shortest isoform, VEGF120, lacks this highly basic domain and is freely diffusible upon secretion. Although the HBD has been attributed significant relevance to VEGF-A biology, the molecular determinants of the heparin-binding site are unknown. We used site-directed mutagenesis to identify amino acid residues that are critical for heparin binding activity of the VEGF164 HBD. We focused on basic residues and found Arg-13, Arg-14, and Arg-49 to be critical for heparin binding and interaction with extracellular matrix in tissue samples. We also examined the cellular and biochemical consequences of abolishing heparin-binding function, measuring the ability of the mutants to interact with VEGF receptors, induce endothelial cell gene expression, and trigger microvessel outgrowth. Induction of tissue factor expression, vessel outgrowth, and binding to VEGFR2 were unaffected by the HBD mutations. In contrast, the HBD mutants showed slightly decreased binding to the NRP1 (neuropilin-1) receptor, and analyses suggested the heparin and NRP1 binding sites to be distinct but overlapping. Finally, mutations that affect the heparin binding activity also led to an unexpected reduction in the affinity of VEGF164 binding specifically to VEGFR1. This finding provides a potential basis for previous observations suggesting enhanced potency of VEGF164 versus VEGF120 in VEGFR1-mediated signaling in inflammatory cells.
Links	PubMed Online version:10.1074/jbc.M700319200
Keywords	Amino Acid Sequence; Animals; Endothelial Cells/cytology; Endothelial Cells/metabolism; Gene Expression Regulation; Heparin/chemistry; Humans; Mice; Molecular Sequence Data; Mutation; Protein Structure, Secondary; Protein Structure, Tertiary; Sequence Homology, Amino Acid; Swine; Vascular Endothelial Growth Factor A/chemistry; Vascular Endothelial Growth Factor A/genetics; Vascular Endothelial Growth Factor A/physiology
edit table

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
MOUSE:VEGFA	acts_upstream_of_or_within	GO:0001525: angiogenesis	ECO:0000314: direct assay evidence used in manual assertion		P		Seeded From UniProt	complete
MOUSE:VEGFA	enables	GO:0005102: signaling receptor binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
MOUSE:VEGFA	enables	GO:0008201: heparin binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
MOUSE:VEGFA	enables	GO:0043183: vascular endothelial growth factor receptor 1 binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
MOUSE:VEGFA	enables	GO:0043184: vascular endothelial growth factor receptor 2 binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete
MOUSE:VEGFA	enables	GO:0050840: extracellular matrix binding	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete

References

See Help:References for how to manage references in GONUTS.