Circulation Research. 2002;90:697-702
Published online before print February 28, 2002, doi: 10.1161/01.RES.0000014227.76102.5D
Published online before print February 28, 2002, doi: 10.1161/01.RES.0000014227.76102.5D
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© 2002 American Heart Association, Inc.
Molecular Medicine |
A Dominant-Negative p65 PAK Peptide Inhibits Angiogenesis
From the Departments of Vascular Biology (W.B.K., D.A.C., N.A., M.A.S.) and Immunology (J.H., D.A.C.), The Scripps Research Institute, La Jolla; and the Department of Cardiovascular Research (S.Y., M.E.G.), Genentech, South San Francisco, Calif.
Correspondence to Martin Alexander Schwartz, Dept of Vascular Biology, The Scripps Research Institute, 10550 North Torrey Pines Rd, La Jolla, CA 92037. E-mail schwartz{at}scripps.edu
PAK1 is a protein kinase downstream of the small GTPases Rac and Cdc42 that previous work has implicated in endothelial cell migration via modulation of cell contraction. The first proline-rich region of PAK that binds to an SH3 domain from the adapter protein NCK was responsible for these dominant-negative effects. To test the role of PAK in angiogenesis, we prepared a peptide in which the proline-rich region was fused to the polybasic sequence from the HIV Tat protein to facilitate entry into cells. We show that the short peptide selectively binds NCK, whereas a mutant peptide does not. Treatment of cells with the PAK peptide but not the control peptide disrupts localization of PAK. This peptide specifically inhibited endothelial cell migration and contractility similarly to full-length dominant-negative PAK. In an in vitro tube-forming assay, the PAK peptide specifically blocked formation of multicellular networks. In an in vivo chick chorioallantoic membrane assay, the PAK peptide specifically blocked angiogenesis. These results, therefore, suggest a role for PAK in angiogenesis.
Key Words: endothelial cells • p21-activated kinase • cell contractility • endothelial tube-forming assay
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