Mechanical Regulation of Actin Binding Proteins

项目摘要 派：丹尼尔·弗莱彻博士。 NIH R01行政副刊 项目名称：肌动蛋白结合蛋白的机械调节 项目期限：2019年7月25日-2023年4月30日 肌动蛋白细胞骨架由细丝的空间组织结构组成，这些结构在结构上是不同的，并且 由300多种不同的肌动蛋白结合蛋白组成。肌动蛋白网络在细胞中的关键作用之一是 产生、传递和抵抗物理力，但肌动蛋白细丝携带的力是否影响 肌动蛋白结合蛋白的结合还不是很清楚。该项目正在研究串联钙蛋白的能力- 同源结构域(CH1-CH2)以张力依赖的方式结合肌动蛋白细丝，并将探索 通过一系列体外和活细胞实验研究张力依赖结合的功能影响。所请求的 FRET-Flim连接将提供一种方法来检验我们的假设，即肌动蛋白的张力依赖变化 细丝可以通过解除结合界面上的空间位阻来改变蛋白质结合亲和力。结果是 这项工作将是对力量如何组织肌动蛋白网络和影响肌动蛋白功能的新理解。 无论是健康细胞还是患病细胞。

Project Summary PI: Daniel Fletcher, Ph.D. NIH R01 Administrative Supplement Project Title: Mechanical Regulation of Actin Binding Proteins Project Period: 7/25/2019 – 4/30/2023 The actin cytoskeleton consists of spatially organized structures of filaments that are architecturally distinct and formed from over 300 different actin-binding proteins. One of the key roles of actin networks in cells is to generate, transmit, and resist physical forces, but whether the forces carried by actin filaments influence the binding of actin-binding proteins is not well understood. This project is investigating the ability of tandem calponin- homology domains (CH1-CH2) to bind actin filaments in a tension-dependent manner, and it will explore the functional impact of tension-dependent binding through a set of in vitro and live cell experiments. The requested FRET-FLIM attachment will provide a way to test our hypothesis that tension-dependent changes in actin filaments can alter protein binding affinity by relieving the steric inhibition at the binding interface. The outcome of this work will be a new understanding of how force can organize actin networks and influence the function of both healthy and diseased cells.