PDF(Pubmed)
Abstract：
Cellular mechanotransduction, a process central to cell biology, embryogenesis, adult physiology and multiple diseases, is thought to be mediated by force-driven changes in protein conformation that control protein function. However, methods to study proteins under defined mechanical loads on a biochemical scale are lacking. We report the development of a DNA based device in which the transition between single-stranded and double-stranded DNA applies tension to an attached protein. Using a fragment of the talin rod domain as a test case, negative-stain electron microscopy reveals programmable extension while pull down assays show tension-induced binding to two ligands, ARPC5L and vinculin, known to bind to cryptic sites inside the talin structure. These results demonstrate the utility of the DNA clamp for biochemical studies and potential structural analysis.
摘要：
细胞机械转导,细胞生物学的核心过程，胚胎发生,成人生理学和多种疾病，被认为是由控制蛋白质功能的蛋白质构象的力驱动变化介导的。然而,缺乏在生化规模上研究定义的机械负荷下的蛋白质的方法。我们报告了一种基于DNA的装置的开发，其中单链和双链DNA之间的过渡将张力施加到附着的蛋白质上。使用塔林棒域的片段作为测试用例，阴性染色电子显微镜显示可编程的延伸，而下拉测定显示张力诱导的结合两个配体，ARPC5L和vinculin，已知与塔林结构内的隐蔽位点结合。这些结果证明了DNA钳在生化研究和潜在结构分析中的实用性。
