Possible role of protein kinase C in the regulation of intracellular stability of focal adhesion kinase in mouse 3T3 cells

FEBS Letters

Fujimoto, Jiro³  Yaoi, Yoshihito¹  Soga, Hisae¹  Takenoshita, Seiichi²  Yokota, Jun¹  Nagamachi, Yukio²  Yamamoto, Tadashi³  Hatai, Mika¹  Mogi, Akira¹ 

[1] Biology Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104, Japan;Department of 1st Surgery, Gunma University School of Medicine, 3-39-15 Showa-machi, Maebashi-shi, Gunma 371, Japan;Department of Oncology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108, Japan

关键词: Calphostin C;    Focal adhesion kinase;    FAK;    Protein kinase C;    Fibronectin;    Phosphorylation;    FAK;    pp125125;    focal adhesion kinase;    2;    5-MeC;    methyl-2;    5-dihydroxycinnamate;    PKC;    protein kinase C;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(95)01014-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Effects of various types of protein kinase inhibitor on the adhesion and spreading of BALB/c mouse 3T3 cells and on the phosphorylation and stability of focal adhesion kinase (FAK) in the cells were studied. Inhibitors of protein tyrosine kinases, methyl 2,5-dihydroxycinnamate and herbimycin A, inhibited tyrosine-phosphorylation of FAK and the adhesion of 3T3 cells to fibronectin. Among inhibitors of serine/threonine kinases tested, calphostin C, a specific inhibitor of protein kinase C, inhibited cell spreading rather than cell adhesion, and it induced the decrease of intracellular FAK within 30 min. Inhibitors of tyrosine kinase, A kinase, G kinase, and myosin light chain kinase did not induce such a rapid and specific decrease of FAK. When calphostin C (20 μM) was added to sub-confluent monolayer cultures, serine-phosphorylation of FAK was inhibited by 67% within 2 h, and decrease in the amount of FAK and rounding up of the cells began after 4 h. Label-chase experiments indicated that about 60% of ³⁵S-labeled FAK degraded within 1–2 h after addition of calphostin C to monolayer cultures. These results indicated that serine-phosphorylation of FAK induced by protein kinase C was important in the regulation of metabolic stability of FAK.

【 授权许可】

Unknown