Talin and signaling through integrins.
Talin and signaling through integrins.
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DOI：
10.1007/978-1-61779-166-6_20
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Calderwood DA
中科院分区：
文献类型：
--
作者：
Bouaouina M;Harburger DS;Calderwood DA
整合素粘附受体对于多细胞动物的发育和功能至关重要。整合素介导细胞与细胞外基质和邻近细胞上的反受体的粘附，并且整合素结合细胞外配体的能力响应于作用于整合素亚基的短胞质尾部的细胞内信号而受到调节。整合素激活，即整合素受体从低亲和力快速转变为高亲和力，需要踝蛋白与整合素β尾部结合，一旦结合，踝蛋白就提供了从激活的整合素到肌动蛋白细胞骨架的连接。多种实验方法有助于目前了解踝蛋白在整合素信号传导中的重要性。在这里，我们描述了两种对 talin 研究至关重要的方法；一种生化测定，可以表征整合素细胞质尾部和踝蛋白之间的相互作用，以及荧光激活细胞分选程序，以评估表达踝蛋白结构域、突变体、显性失活结构或 shRNA 的培养细胞中的整合素激活。
Integrin adhesion receptors are essential for development and functioning of multi-cellular animals. Integrins mediate cell adhesion to the extracellular matrix and to counter-receptors on adjacent cells and the ability of integrins to bind extracellular ligands is regulated in response to intracellular signals that act on the short cytoplasmic tails of integrin subunits. Integrin activation, the rapid conversion of integrin receptors from low to high affinity, requires binding of talin to integrin β tails and, once bound, talin provides a connection from activated integrins to the actin cytoskeleton. A wide range of experimental approaches have contributed to the current understanding of the importance of talin in integrin signaling. Here we describe two methods that have been central to our investigations of talin; a biochemical assay that has allowed characterization of interactions between integrin cytoplasmic tails and talin and a fluorescent activated cell sorting procedure to assess integrin activation in cultured cells expressing talin domains, mutants, dominant negative constructs, or shRNA.