Measurements of Myosin-II Motor Activity During Cytokinesis in Fission Yeast.
Measurements of Myosin-II Motor Activity During Cytokinesis in Fission Yeast.
复制标题
裂殖酵母细胞分裂过程中肌球蛋白-II 运动活性的测量。
DOI：
10.1007/978-1-4939-3145-3_11
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Lord,Matthew
中科院分区：
文献类型：
--
作者：
Tang,Qing;Pollard,LutherW;Lord,Matthew
裂变酵母肌凝蛋白ii （Myo2p）是一种关键的基于肌动蛋白的运动蛋白，在细胞分裂过程中驱动肌动蛋白环的组装和收缩。我们详细介绍了测量Myo2p运动功能的三种不同方法。肌动蛋白激活的atp酶在批量检测中提供肌动球蛋白atp酶运动活性的读数；肌动蛋白丝运动分析揭示了肌球蛋白驱动的肌动蛋白丝滑动的速度和效率（当马达被锚定时）；肌凝蛋白头运动分析揭示了肌凝蛋白群沿肌动蛋白丝移动的速度和效率（当肌动蛋白被锚定时）。总的来说，这些方法使我们能够将裂变酵母常见的标准体内方法与体外生化方法相结合，以更多地了解肌球蛋白- ii在细胞分裂过程中的机制作用。
Fission yeast myosin-II (Myo2p) represents the critical actin-based motor protein that drives actomyosin ring assembly and constriction during cytokinesis. We detail three different methods to measure Myo2p motor function. Actin-activated ATPases provide a readout of actomyosin ATPase motor activity in a bulk assay; actin filament motility assays reveal the speed and efficiency of myosin-driven actin filament gliding (when motors are anchored); myosin-bead motility assays reveal the speed and efficiency of myosin ensembles traveling along actin filaments (when actin is anchored). Collectively, these methods allow us to combine the standard in vivo approaches common to fission yeast with in vitro biochemical methods to learn more about the mechanistic action of myosin-II during cytokinesis.