Mechanical Regulation of Actin Binding Proteins

项目摘要 PI：丹尼尔弗莱彻，博士 NIH R 01行政补充 项目名称：肌动蛋白结合蛋白的机械调节 计划日期：2019年7月25日至2023年4月30日 肌动蛋白细胞骨架由空间上有组织的细丝结构组成，这些结构在结构上是不同的， 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，但是肌动蛋白丝携带的力是否影响了细胞的生长， 肌动蛋白结合蛋白的结合还没有被很好地理解。这个项目正在研究串联的钙调蛋白的能力- 同源结构域（CH 1-CH 2）以张力依赖的方式结合肌动蛋白丝，它将探索 通过一组体外和活细胞实验，研究张力依赖性结合的功能影响。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性变化 细丝可以通过减轻结合界面处的空间抑制来改变蛋白质结合亲和力。成果 这项工作将是一个新的理解力如何组织网络和影响功能的行动 健康和患病的细胞。

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.