ADAMTS1/METH1 inhibits endothelial cell proliferation by direct binding and sequestration of VEGF165.
ADAMTS1 is a metalloprotease previously shown to inhibit angiogenesis in a variety of in vitro and in vivo assays. In the present study, we demonstrate that ADAMTS1 significantly blocks VEGFR2 phosphorylation with consequent suppression of endothelial cell proliferation. The effect on VEGFR2 function was due to direct binding and sequestration of VEGF165 by ADAMTS1. Binding was confirmed by co-immunoprecipitation and cross-linking analysis. Inhibition of VEGF function was reversible, as active VEGF could be recovered from the complex. The interaction required the heparin-binding domain of the growth factor, because VEGF121 failed to bind to ADAMTS1. Structure/function analysis with independent ADAMTS1 domains indicated that binding to VEGF165 was mediated by the carboxyl-terminal (CT) region. ADAMTS1 and VEGF165 were also found in association in tumor extracts. These findings provide a mechanism for the anti-angiogenic activity of ADAMTS1 and describe a novel modulator of VEGF bioavailability.
Luque
Alfonso
A
Department of Molecular, University of California, Los Angeles, California 90095, USA.
Carpizo
Darren R
DR
Iruela-Arispe
M Luisa
ML
D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; D000818:Animals; D001682:Biological Availability; D002417:Cattle; D002455:Cell Division; D019483:Disintegrins / Q000378:metabolism / Q000502:physiology*; D016228:Endothelial Growth Factors / Q000378:metabolism* / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism* / Q000494:pharmacology; D008222:Lymphokines / Q000378:metabolism* / Q000494:pharmacology; D008666:Metalloendopeptidases / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000473:pathology; D018919:Neovascularization, Physiologic; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014407:Tumor Cells, Cultured; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D042442:Vascular Endothelial Growth Factors
D019483:Disintegrins; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D040301:Vascular Endothelial Growth Factor Receptor-2; D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; C103859:ADAMTS1 protein, human; C486323:Adamts1 protein, mouse; D008666:Metalloendopeptidases