Identification of a novel protein complex containing annexin VI, Fyn, Pyk2, and the p120 GAP C2 domain

Abstract

p120 GAP (RasGAP) has been proposed to function as both an inhibitor and effector of Ras. Previously we have shown that RasGAP contains a C2 domain which mediates both Ca 2+-dependent membrane association and protein–protein interactions. Specifically, three proteins have been isolated in a complex with the C2 domain of RasGAP; these are the Ca 2+-dependent lipid binding protein annexin VI (p70) and two previously unidentified proteins, p55 and p120. Here we provide evidence that p55 is the Src family kinase Fyn and p120 is the focal adhesion kinase family member Pyk2. In addition, in vitro binding assays indicate that Fyn, but not Pyk2 binds directly to annexin VI. Finally, co-immunoprecipitation studies in Rat-1 fibroblasts confirm that Fyn, Pyk2, annexin VI and RasGAP can form a protein complex in mammalian cells.