Speeding up kinesin-driven microtubule gliding in vitro by variation of cofactor composition and physicochemical parameters
Speeding up kinesin-driven microtubule gliding in vitro by variation of cofactor composition and physicochemical parameters
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DOI：
10.1006/cbir.1999.0515
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发表时间：
2000-01-01
影响因子：
3.9
通讯作者：
Unger, E
中科院分区：
文献类型：
--
作者：
Böhm, KJ;Stracke, R;Unger, E
到目前为止，在体外和细胞内测量的驱动蛋白依赖性微管运动的速度之间存在差异。通过更改ATP，MG2+和动力蛋白浓度，pH和离子强度，我们试图找到有利于微管滑行的条件。对于猪脑驱动蛋白，我们发现提高摩尔MG2+/ATP比率可以显着提高滑行速度。温度升高或减少与玻璃表面结合的驱动蛋白分子的数量也变得更快。通过有利因素的组合（2.5 mm ATP，12.5 mm mg2 +，37 +，37级C，37度，37毫米，37毫米，平均滑行速度（1.8 mU m/s +/- 0.09 mu m/s），接近实行速度的速度达到了实行速度。 450个驱动蛋白分子/MU M（2））。 （c）2000学术出版社
So far, there has been a discrepancy between the velocities of kinesin-dependent microtubule motility measured in vitro and within cells. By changing ATP, Mg2+, and kinesin concentrations, pH and ionic strength, we tried to find conditions that favour microtubule gliding across kinesin-covered glass surfaces. For porcine brain kinesin, we found that raising the molar Mg2+/ATP ratio can substantially elevate gliding velocity. Gliding became also faster after temperature elevation or lowering the number of kinesin molecules bound to the glass surface. The highest mean gliding velocity (1.8 mu m/s +/- 0.09 mu m/s), approaching velocities measured for anterograde transport in vivo, was achieved by combination of favourable factors (2.5 mM ATP, 12.5 mM Mg2+, 37 degrees C, 450 kinesin molecules/mu m(2)). (C) 2000 Academic Press