Spudich James A, Rice Sarah E, Rock Ronald S, Purcell Thomas J, Warrick Hans M
Cold Spring Harb Protoc. 2011 Nov 1;2011(11):1305-18. doi: 10.1101/pdb.top066662.
In vitro motility assays enabled the analysis of coupling between ATP hydrolysis and movement of myosin along actin filaments or kinesin along microtubules. Single-molecule assays using laser trapping have been used to obtain more detailed information about kinesins, myosins, and processive DNA enzymes. The combination of in vitro motility assays with laser-trap measurements has revealed detailed dynamic structural changes associated with the ATPase cycle. This article describes the use of optical traps to study processive and nonprocessive molecular motor proteins, focusing on the design of the instrument and the assays to characterize motility.
体外运动分析能够分析ATP水解与肌球蛋白沿肌动蛋白丝的运动或驱动蛋白沿微管的运动之间的偶联。使用激光捕获的单分子分析已用于获取有关驱动蛋白、肌球蛋白和进行性DNA酶的更详细信息。体外运动分析与激光捕获测量的结合揭示了与ATP酶循环相关的详细动态结构变化。本文描述了使用光镊研究进行性和非进行性分子运动蛋白，重点是仪器的设计和表征运动性的分析方法。
