Cooperative Role of Caveolin-1 and C-Terminal Src Kinase Binding Protein in C-Terminal Src Kinase-Mediated Negative Regulation of c-Src

被引:34
作者
Place, Aaron T. [1 ]
Chen, Zhenlong [1 ]
Bakhshi, Farnaz R. [1 ]
Liu, Guoquan
O'Bryan, John P. [1 ]
Minshall, Richard D. [1 ,2 ,3 ]
机构
[1] Univ Illinois, Dept Pharmacol, Chicago, IL 60612 USA
[2] Univ Illinois, Dept Anesthesiol, Chicago, IL 60612 USA
[3] Univ Illinois, Ctr Lung & Vasc Biol, Chicago, IL 60612 USA
基金
美国国家卫生研究院;
关键词
FAMILY TYROSINE KINASES; IN-VIVO; ADAPTER PROTEIN; CSK GENE; PHOSPHORYLATION; CBP; ACTIVATION; ENDOTHELIUM; ENDOCYTOSIS; METASTASIS;
D O I
10.1124/mol.111.073957
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
In the present study, we assessed the cooperative roles of C-terminal Src kinase (Csk) binding protein (Cbp) and Caveolin-1 (Cav-1) in the mechanism of Src family tyrosine kinase (SFK) inhibition by Csk. SFKs are inactivated by phosphorylation of their C-terminal tyrosine by Csk. Whereas SFKs are membrane-associated, Csk is a cytoplasmic protein and therefore requires membrane adaptors such as Cbp or Cav-1 for recruitment to the plasma membrane to mediate SFK inhibition. To determine the specific role of Cav-1 and Cbp in SFK inhibition, we measured c-Src activity in the absence of each membrane adaptor. It is noteworthy that in lungs and fibroblasts from Cav-1(-/-) mice, we observed increased expression of Cbp compared with wild-type (WT) controls. However, both c-Src activity and Csk localization at the membrane were similar between Cav-1(-/-) fibroblasts and WT cells. Likewise, Cbp depletion by small interfering RNA (siRNA) treatment of WT cells had no effect on basal c-Src activity, but it increased the phosphorylation state of Cav-1. Immunoprecipitation then confirmed increased association of Csk with phosphomimicking Cav-1. Knockdown of Cbp by siRNA in Cav-1(-/-) cells revealed increased basal c-Src activity, and re-expression of WT Cav-1 in the same cells reduced basal c-Src activity. Taken together, these results indicate that Cav-1 and Cbp cooperatively regulate c-Src activity by recruiting Csk to the membrane where it phosphorylates c-Src inhibitory tyrosine 529. Furthermore, when either Cav-1 or Cbp expression is reduced or absent, there is a compensatory increase in the phosphorylation state or expression level of the other membrane-associated Csk adaptor to maintain SFK inhibition.
引用
收藏
页码:665 / 672
页数:8
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