Bakolitsa Constantina, Cohen Daniel M, Bankston Laurie A, Bobkov Andrey A, Cadwell Gregory W, Jennings Lisa, Critchley David R, Craig Susan W, Liddington Robert C
Program on Cell Adhesion, The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, California 92037, USA.
Nature. 2004 Jul 29;430(6999):583-6. doi: 10.1038/nature02610. Epub 2004 Jun 13.
Vinculin is a highly conserved intracellular protein with a crucial role in the maintenance and regulation of cell adhesion and migration. In the cytosol, vinculin adopts a default autoinhibited conformation. On recruitment to cell-cell and cell-matrix adherens-type junctions, vinculin becomes activated and mediates various protein-protein interactions that regulate the links between F-actin and the cadherin and integrin families of cell-adhesion molecules. Here we describe the crystal structure of the full-length vinculin molecule (1,066 amino acids), which shows a five-domain autoinhibited conformation in which the carboxy-terminal tail domain is held pincer-like by the vinculin head, and ligand binding is regulated both sterically and allosterically. We show that conformational changes in the head, tail and proline-rich domains are linked structurally and thermodynamically, and propose a combinatorial pathway to activation that ensures that vinculin is activated only at sites of cell adhesion when two or more of its binding partners are brought into apposition.
纽蛋白是一种高度保守的细胞内蛋白质,在维持和调节细胞黏附与迁移中起关键作用。在胞质溶胶中,纽蛋白呈现默认的自抑制构象。当被招募到细胞间和细胞-基质黏附型连接时,纽蛋白被激活并介导各种蛋白质-蛋白质相互作用,这些相互作用调节F-肌动蛋白与细胞黏附分子的钙黏蛋白和整合素家族之间的连接。在此,我们描述了全长纽蛋白分子(1066个氨基酸)的晶体结构,其显示出一种五结构域自抑制构象,其中羧基末端尾部结构域被纽蛋白头部以钳状方式夹住,并且配体结合在空间上和别构上均受到调节。我们表明,头部、尾部和富含脯氨酸结构域的构象变化在结构和热力学上是相关联的,并提出了一种激活的组合途径,该途径确保纽蛋白仅在其两个或更多结合伴侣并列时的细胞黏附位点处被激活。