raf kinase inhibitory protein protects cells against locostatin-mediated inhibition of migration英国皇家空军激酶抑制蛋白质保护细胞免受locostatin-mediated抑制迁移.pdfVIP

Raf Kinase Inhibitory Protein Protects Cells against Locostatin-Mediated Inhibition of Migration 1 1 1,2 3 1 1 Anne N. Shemon , Eva M. Eves , Matthew C. Clark , Gary Heil , Alexey Granovsky , Lingchun Zeng , Akira Imamoto1, Shohei Koide3.*, Marsha Rich Rosner1,2.* 1 Ben May Department for Cancer Research, University of Chicago, Chicago, Illinois, United States of America, 2 Department of Neurobiology, Pharmacology and Physiology, University of Chicago, Chicago, Illinois, United States of America, 3 Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, Illinois, United States of America Abstract Background: Raf Kinase Inhibitory Protein (RKIP, also PEBP1), a member of the Phosphatidylethanolamine Binding Protein family, negatively regulates growth factor signaling by the Raf/MAP kinase pathway. Since an organic compound, locostatin, was reported to bind RKIP and inhibit cell migration by a Raf-dependent mechanism, we addressed the role of RKIP in locostatin function. Methods/Findings: We analyzed locostatin interaction with RKIP and examined the biological consequences of locostatin binding on RKIP function. NMR studies show that a locostatin precursor binds to the conserved phosphatidylethanolamine binding pocket of RKIP. However, drug binding to the pocket does not prevent RKIP association with its inhibitory target, Raf-1, nor affect RKIP phosphorylation by Protein Kinase C at a regulatory site. Similarly, exposure of wild type, RKIP- depleted HeLa cells or RKIP-deficient (RKIP2/ 2) mouse embryonic fibroblasts (MEFs) to locostatin has no effect on MAP kinase activation. Locostatin treatment of wild t