In vitro assays of processive myosin motors
In vitro assays of processive myosin motors
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DOI：
10.1006/meth.2000.1089
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发表时间：
2000-12-01
期刊：
影响因子：
4.8
通讯作者：
Spudich, JA
中科院分区：
文献类型：
--
作者：
Rock, RS;Rief, M;Spudich, JA
肌凝蛋白V是一种基于动作蛋白的运动蛋白，被认为参与囊泡运输。由于这种运动的性质可能与肌球蛋白II不同，我们使用滑动丝和光学陷阱分析的组合来观察高分辨率的单分子，研究了肌球蛋白v驱动的运动。结果清楚地表明，脑肌球蛋白V是一种高效的进程马达。体外运动测定法肌球蛋白V密度显示明显的单分子支持运动。在肌球蛋白v驱动运动的光学捕获实验中也观察到渐进步进。这里描述了用于证明肌凝蛋白V的处理能力的方法。这些方法包括密度依赖的分析，消除了多个马达聚集或机会共定位的可能性，这些马达是造成明显的单分子运动的原因。这种检测方法将是鉴别其他进展类肌凝蛋白的有用工具。(C) 2000年学术出版社。
Myosin V is an actin-based motor thought to be involved in vesicle transport. Since the properties of such a motor may be expected to differ from those of muscle myosin II, we have examined myosin V-driven movement using a combination of gliding filament and optical trap assays to observe single molecules with high resolution. The results clearly demonstrate that brain: myosin V is a highly efficient processive motor. In vitro motility assays at law myosin V densities reveal apparent single-molecule supported movement. Processive stepping was also observed in optical trapping assays of myosin V-driven motion. Here the methods that were used to demonstrate the processivity of myosin V are described. These methods include density-dependent assays that eliminate the possibility of aggregation or chance colocalization of multiple motors being responsible for apparent single-molecule motility. Such assays will be useful tools far identifying other processive classes of myosins. (C) 2000 Academic Press.