Suppression of Pyk2 kinase and cellular activities by FIP200.
Suppression of Pyk2 kinase and cellular activities by FIP200.
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DOI：
10.1083/jcb.149.2.423
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发表时间：
2000-04-17
期刊：
影响因子：
--
通讯作者：
Guan JL
中科院分区：
文献类型：
--
作者：
Ueda H;Abbi S;Zheng C;Guan JL
富含脯氨酸的酪氨酸激酶2 （Pyk2）是一种细胞质酪氨酸激酶，在多种细胞内信号通路中发挥作用。我们报道了利用酵母双杂交筛选鉴定了一种新的pyk2相互作用蛋白FIP200 （FAK家族激酶相互作用蛋白200 kD）。体外结合实验和共免疫沉淀证实了FIP200与Pyk2的关联，类似的实验也表明FIP200与FAK结合。然而，免疫荧光染色显示FIP200主要定位于细胞质中。FIP200结合Pyk2的激酶结构域并抑制其激酶活性。FIP200还抑制SYF细胞中分离的Pyk2 （Src、Yes和Fyn表达不足）和缺乏Src结合位点的Pyk2突变体的激酶活性，表明它直接调节Pyk2激酶，而不是影响相关的Src家族激酶。与体外抑制作用一致，FIP200抑制了Pyk2的激活和Pyk2诱导的完整细胞凋亡，这与FIP200与Pyk2的结合有关。最后，几种生物刺激对Pyk2的激活与内源性FIP200 - Pyk2复合物的解离有关，这进一步支持了FIP200在完整细胞中对Pyk2的抑制作用。总之，这些结果表明FIP200通过结合其激酶结构域作为Pyk2的抑制剂起作用。
Proline-rich tyrosine kinase 2 (Pyk2) is a cytoplasmic tyrosine kinase implicated to play a role in several intracellular signaling pathways. We report the identification of a novel Pyk2-interacting protein designated FIP200 (FAK family kinase–interacting protein of 200 kD) by using a yeast two-hybrid screen. In vitro binding assays and coimmunoprecipitation confirmed association of FIP200 with Pyk2, and similar assays also showed FIP200 binding to FAK. However, immunofluorescent staining indicated that FIP200 was predominantly localized in the cytoplasm. FIP200 bound to the kinase domain of Pyk2 and inhibited its kinase activity in in vitro kinase assays. FIP200 also inhibited the kinase activity of the Pyk2 isolated from SYF cells (deficient in Src, Yes, and Fyn expression) and the Pyk2 mutant lacking binding site for Src, suggesting that it regulated Pyk2 kinase directly rather than affecting the associated Src family kinases. Consistent with its inhibitory effect in vitro, FIP200 inhibited activation of Pyk2 and Pyk2-induced apoptosis in intact cells, which correlated with its binding to Pyk2. Finally, activation of Pyk2 by several biological stimuli correlated with the dissociation of endogenous FIP200–Pyk2 complex, which provided further support for inhibition of Pyk2 by FIP200 in intact cells. Together, these results suggest that FIP200 functions as an inhibitor of Pyk2 via binding to its kinase domain.
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DOI：
10.1093/dnares/3.5.321
发表时间：
1996-10-31
期刊：
DNA research : an international journal for rapid publication of reports on genes and genomes
影响因子：
--
作者：
Nagase, T;Seki, N;Nomura, N
通讯作者：
Nomura, N
影响因子：
4.8
作者：
Brinson, AE;Harding, T;Graves, LM
通讯作者：
Graves, LM
影响因子：
11.4
作者：
Sieg, DJ;Ilic, D;Schlaepfer, DD
通讯作者：
Schlaepfer, DD
影响因子：
4.8
作者：
Li, X;Earp, HS
通讯作者：
Earp, HS
影响因子：
64.8
作者：
LEV, S;MORENO, H;SCHLESSINGER, J
通讯作者：
SCHLESSINGER, J