Quantifying Strain-Sensing Protein Recruitment During Stress Fiber Repair.
Quantifying Strain-Sensing Protein Recruitment During Stress Fiber Repair.
复制标题
量化应力纤维修复过程中应变感应蛋白的募集。
DOI：
10.1007/978-1-0716-2851-5_11
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Oakes,PatrickW
中科院分区：
文献类型：
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作者：
Seetharaman,Shailaja;Sala,Stefano;Gardel,MargaretL;Oakes,PatrickW
已经鉴定出一个蛋白质家族，可以识别应力纤维中受损、紧张的肌动蛋白丝。这些蛋白质通常被称为应变或力感应，并触发下游信号机制，从而促进这些应变位点的修复。在这里，我们描述了一种使用高分辨率显微镜来筛选和量化这些应力纤维应变位点的蛋白质的机械敏感募集的方法。使用空间控制的紫外光照射诱导应变位点，以局部损伤肌动蛋白应力纤维。然后，可以将潜在应变感应蛋白的招募与（Blanchoin, Physiol Rev 94, 235–263, 2014）已知对照（例如 zyxin-GFP）或（Hoffman, Mol Biol Cell 23, 1846–1859, 2012）预损坏的应力纤维蛋白分布进行比较。通过这种方法，可以重复地测量和比较应变传感蛋白及其与应力纤维应变位点的动态关联。
A family of proteins have been identified that recognize damaged, strained actin filaments in stress fibers. These proteins are often referred to as strain- or force-sensing and trigger downstream signaling mechanisms that can facilitate repair at these strain sites. Here we describe a method using high-resolution microscopy to screen and quantify the mechanosensitive recruitment of proteins to these stress fiber strain sites. Strain sites are induced using spatially controlled illumination of UV light to locally damage actin stress fibers. Recruitment of potential strain-sensing proteins can then either be compared to (Blanchoin, Physiol Rev 94, 235–263, 2014) a known control (e.g., zyxin-GFP) or (Hoffman, Mol Biol Cell 23, 1846–1859, 2012) the pre-damaged stress fiber protein distribution. With this method, strain-sensing proteins and their dynamic association with stress fiber strain sites can be reproducibly measured and compared.