Identification of a novel protein complex containing annexin VI, Fyn, Pyk2, and the p120GAP C2 domain

FEBS Letters

Davis, Alison J¹  Gawler, Debra J¹  Chow, Andrew¹ 

[1] School of Biomedical Sciences, University of Leeds, Woodhouse Lane, Leeds LS2 9JT, UK

关键词: Conserved region 2;    GTPase activating protein;    Ras;    Annexin;    Ca2+;    GAP;    GTPase activating protein;    C2;    conserved region 2;    GST;    glutathione S-transferase;    Pyk2;    proline rich tyrosine kinase 2;    EGTA;    ethylene glycol-bis(β-amino ethyl ether)-N′;    N′;    N′;    N′-tetraacetic acid;    SDS–PAGE;    sodium dodecyl sulphate–polyacrylamide gel electrophoresis;    PBS;    phosphate buffered saline;    HRP;    horseradish peroxidase;    ECL;    enhanced chemiluminescence;   

DOI  :  10.1016/S0014-5793(00)01252-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 Ca²⁺-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 Ca²⁺-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.

【 授权许可】

Unknown