Byron Adam, Askari Janet A, Humphries Jonathan D, Jacquemet Guillaume, Koper Ewa J, Warwood Stacey, Choi Colin K, Stroud Matthew J, Chen Christopher S, Knight David, Humphries Martin J
Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
Biological Mass Spectrometry Core Facility, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, UK.
Nat Commun. 2015 Jan 22;6:6135. doi: 10.1038/ncomms7135.
Integrin activation, which is regulated by allosteric changes in receptor conformation, enables cellular responses to the chemical, mechanical and topological features of the extracellular microenvironment. A global view of how activation state converts the molecular composition of the region proximal to integrins into functional readouts is, however, lacking. Here, using conformation-specific monoclonal antibodies, we report the isolation of integrin activation state-dependent complexes and their characterization by mass spectrometry. Quantitative comparisons, integrating network, clustering, pathway and image analyses, define multiple functional protein modules enriched in a conformation-specific manner. Notably, active integrin complexes are specifically enriched for proteins associated with microtubule-based functions. Visualization of microtubules on micropatterned surfaces and live cell imaging demonstrate that active integrins establish an environment that stabilizes microtubules at the cell periphery. These data provide a resource for the interrogation of the global molecular connections that link integrin activation to adhesion signalling.
整合素激活由受体构象的变构变化调节，使细胞能够对细胞外微环境的化学、机械和拓扑特征做出反应。然而，目前尚缺乏关于激活状态如何将整合素近端区域的分子组成转化为功能读数的整体观点。在这里，我们使用构象特异性单克隆抗体，报告了整合素激活状态依赖性复合物的分离及其质谱表征。通过整合网络、聚类、通路和图像分析进行的定量比较，定义了以构象特异性方式富集的多个功能蛋白模块。值得注意的是，活性整合素复合物特别富集与基于微管功能相关的蛋白质。在微图案化表面上对微管进行可视化以及活细胞成像表明，活性整合素建立了一个在细胞周边稳定微管的环境。这些数据为探究将整合素激活与粘附信号联系起来的整体分子连接提供了资源。
