Turn-On Protein Switches for Controlling Actin Binding in Cells.
Turn-On Protein Switches for Controlling Actin Binding in Cells.
复制标题
打开控制细胞中肌动蛋白结合的蛋白质开关。
DOI：
10.1101/2023.10.26.561921
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Belardi,Brian
中科院分区：
文献类型：
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作者：
Effiong,UnyimeM;Khairandish,Hannah;Ramirez-Velez,Isabela;Wang,Yanran;Belardi,Brian
在一个共享的细胞质中，丝状肌动蛋白（F-actin）在整个细胞体中发挥着许多关键作用。细胞依靠肌动蛋白结合蛋白（ABP）来组织F-肌动蛋白，并将其聚合特性整合到不同的细胞过程中。然而，众多的ABP参与并塑造F-肌动蛋白，使研究单个ABP对细胞活动的影响成为一个重大挑战。此外，没有一种操纵肌动蛋白结合亚细胞的手段，利用F-肌动蛋白细胞骨架合成生物学的目的仍然难以捉摸。在这里，我们描述了一套设计的蛋白质，可控肌动蛋白结合开关工具（CAST），其肌动蛋白结合行为可以控制与外部刺激。开发了响应不同外部输入的CAST，提供了开启动力学的选择，并实现了正交性和多路复用。通过遗传编码，我们发现CAST可以插入到天然蛋白质序列中，以控制局部的F-肌动蛋白结合，并设计成结构来控制细胞和组织的形状和行为。
Within a shared cytoplasm, filamentous actin (F-actin) plays numerous and critical roles across the cell body. Cells rely on actin-binding proteins (ABPs) to organize F-actin and to integrate its polymeric characteristics into diverse cellular processes. Yet, the multitude of ABPs that engage with and shape F-actin make studying a single ABP’s influence on cellular activities a significant challenge. Moreover, without a means of manipulating actin-binding subcellularly, harnessing the F-actin cytoskeleton for synthetic biology purposes remains elusive. Here, we describe a suite of designed proteins, Controllable Actin-binding Switch Tools (CASTs), whose actin-binding behavior can be controlled with external stimuli. CASTs were developed that respond to different external inputs, providing options for turn-on kinetics and enabling orthogonality and multiplexing. Being genetically encoded, we show that CASTs can be inserted into native protein sequences to control F-actin association locally and engineered into structures to control cell and tissue shape and behavior.