Regulation of Adhesion Dynamics by Calpain-mediated Proteolysis of Focal Adhesion Kinase (FAK)
Regulation of Adhesion Dynamics by Calpain-mediated Proteolysis of Focal Adhesion Kinase (FAK)
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DOI：
10.1074/jbc.m109.090746
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发表时间：
2010-04-09
影响因子：
4.8
通讯作者：
Huttenlocher, Anna
中科院分区：
文献类型：
--
作者：
Chan, Keefe T.;Bennin, David A.;Huttenlocher, Anna
粘附的协调和动态调节是细胞迁移所必需的。我们之前已经证明，钙依赖性蛋白酶钙蛋白酶2对塔林1的有限蛋白水解在成纤维细胞的粘附分解中发挥着关键作用（Franco，SJ，Rodgers，M.一、Perrin，B.J.，汉，J.，Bennin，D.一、Critchley，D. R.，和Huttenlocher，A.（2004）Nat. Cell Biol.6，977-983）。然而，鲜为人知的是其他钙蛋白酶底物的粘附动力学的调节的贡献。我们现在提供的证据表明，钙蛋白酶2介导的粘着斑激酶（FAK）的蛋白水解调节运动细胞的粘附动力学。我们绘制了首选的钙蛋白酶切割位点之间的两个C-末端脯氨酸丰富的区域后，Ser-745，在C-末端片段的大小相似的FAK相关的非激酶（FRNK）。我们产生了突变FAK与点突变（V744 G），使FAK耐钙蛋白酶蛋白水解，但保留其他生化特性的FAK。使用延时显微镜，我们表明，绿色荧光蛋白talin 1的动力学在FAK缺陷细胞受损。表达野生型但不耐钙蛋白酶的FAK拯救FAK缺陷细胞中的塔林动力学两者合计，我们的研究结果表明，钙蛋白酶蛋白水解FAK在调节运动细胞的粘附动力学的一个新的作用。
The coordinated and dynamic regulation of adhesions is required for cell migration. We demonstrated previously that limited proteolysis of talin1 by the calcium-dependent protease calpain 2 plays a critical role in adhesion disassembly in fibroblasts (Franco, S.J., Rodgers, M. A., Perrin, B.J., Han, J., Bennin, D. A., Critchley, D. R., and Huttenlocher, A. (2004) Nat. Cell Biol. 6, 977-983). However, little is known about the contribution of other calpain substrates to the regulation of adhesion dynamics. We now provide evidence that calpain 2-mediated proteolysis of focal adhesion kinase (FAK) regulates adhesion dynamics in motile cells. We mapped the preferred calpain cleavage site between the two C-terminal proline-rich regions after Ser-745, resulting in a C-terminal fragment similar in size to the FAK-related non-kinase (FRNK). We generated mutant FAK with a point mutation (V744G) that renders FAK resistant to calpain proteolysis but retains other biochemical properties of FAK. Using time-lapse microscopy, we show that the dynamics of green fluorescent protein-talin1 are impaired in FAK-deficient cells. Expression of wild-type but not calpain-resistant FAK rescues talin dynamics in FAK-deficient cells. Taken together, our findings suggest a novel role for calpain proteolysis of FAK in regulating adhesion dynamics in motile cells.