Characterization of the interaction of Raf‐1 with ras p21 or 14‐3‐3 protein in intact cells

FEBS Letters

Koyama, Shinya¹  Williams, Lewis T.²  Kikuchi, Akira¹ 

[1] Department of Biochemistry, Hiroshima University School of Medicine, Hiroshima 734, Japan;Daiichi Research Center, Cardiovascular Research Institute, University of California, San Francisco, CA 94143-0130, USA

关键词: 14-3-3 Protein;    ras p21;    Raf-1;    COS cell;    Spodoptera grugiperda cell (Sf9 cell);    MAP kinase;    mitogen-activated protein kinase;    HA;    hemagglutinin;    GST;    glutathione-S-transferase;    PCR;    polymerase chain reaction;    Sf9 cells;    Spodoptera frugiperda cells;    IP;    immunoprecipitation;   

DOI  :  10.1016/0014-5793(95)00686-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Several deletion mutants of Raf-1 were expressed with v-ras p21 or 14-3-3 protein in COS-7 cells and Sf9 cells and the interaction of Raf-1 with ras p21 or with 14-3-3 protein in intact cells was examined. Raf(1–135) (residues 1–135) and Raf(1–322) interacted with v-ras p21, but other deletion mutants such as Raf(136–322) or Raf(321–648) did not. Raf(1–322) interacted with 14-3-3 protein much more efficiently than Raf(321–648) did. While Raf(1–135) did not interact with 14-3-3 protein, Raf(136–322) did. These results clearly indicate that Raf-1 simultaneously interacts with both ras p21 and 14-3-3 protein through the distinct binding domains in intact cells.

【 授权许可】

Unknown