Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface.
Tug-of-war of microtubule filaments at the boundary of a kinesin- and dynein-patterned surface.
复制标题
DOI：
10.1038/srep05281
复制
发表时间：
2014-06-13
影响因子：
4.6
通讯作者：
Yokokawa R
中科院分区：
文献类型：
--
作者：
Ikuta J;Kamisetty NK;Shintaku H;Kotera H;Kon T;Yokokawa R
细胞内的货物由多种运动蛋白运输。由于混合极性马达的力平衡，货物可以双向移动，实现生物功能。在这里，我们提出了一种微管滑动试验，用于对动力蛋白和动力蛋白的拔河研究。两个马达组的边界是通过光刻模式金来选择性地将动力蛋白附着在玻璃上，并使用自组装的单层将动力蛋白附着在金表面。由各电机的力-速度关系推导出两个对抗电机数的比值与速度的关系，从而计算出分离力和电机后退速度。尽管这场拔河涉及到100个马达，但数值是为单个分子计算出来的，反映了集体动力蛋白和非集体动力蛋白协同工作时的功能。该分析将用于细胞内运动的详细体外分析，例如，有丝分裂，其中涉及大量具有混合极性的马达。
Intracellular cargo is transported by multiple motor proteins. Because of the force balance of motors with mixed polarities, cargo moves bidirectionally to achieve biological functions. Here, we propose a microtubule gliding assay for a tug-of-war study of kinesin and dynein. A boundary of the two motor groups is created by photolithographically patterning gold to selectively attach kinesin to the glass and dynein to the gold surface using a self-assembled monolayer. The relationship between the ratio of two antagonistic motor numbers and the velocity is derived from a force-velocity relationship for each motor to calculate the detachment force and motor backward velocity. Although the tug-of-war involves >100 motors, values are calculated for a single molecule and reflect the collective dynein and non-collective kinesin functions when they work as a team. This assay would be useful for detailed in vitro analysis of intracellular motility, e.g., mitosis, where a large number of motors with mixed polarities are involved.