Hirakawa E, Higuchi H, Toyoshima Y Y
Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Tokyo 153-8902, Japan.
Proc Natl Acad Sci U S A. 2000 Mar 14;97(6):2533-7. doi: 10.1073/pnas.050585297.
We have analyzed the movement of single 22S dynein molecules from Tetrahymena cilia by using a nanometer measuring system equipped with optical tweezers. Statistical analysis proved that a single molecule of 22S dynein can move processively and develop force at low concentrations of ATP (<20 microM). The maximum force was approximately 4.7 pN, and the force-velocity curve was convex down. During force development, dynein molecules showed stepwise displacement of approximately 8 nm and frequently exhibited backward steps of approximately 8 nm. At higher concentrations of ATP (>/=20 microM) single molecules of 22S dynein were not observed to move processively. Twenty-two S dynein seems to switch over from a processive mode to a nonprocessive mode, sensing a subtle change of ATP concentrations. These observations indicate that the processivity, maximum force, and step size of dynein are similar to those of kinesin, but the ATP concentration-dependence, force-velocity relationship, and backward steps are clearly distinct from kinesin.
我们使用配备光镊的纳米测量系统,分析了来自四膜虫纤毛的单个22S动力蛋白分子的运动。统计分析表明,单个22S动力蛋白分子在低浓度ATP(<20 microM)时能够持续移动并产生力。最大力约为4.7皮牛,力-速度曲线向下凸出。在产生力的过程中,动力蛋白分子显示出约8纳米的逐步位移,并频繁出现约8纳米的向后位移。在较高浓度的ATP(≥20 microM)下,未观察到单个22S动力蛋白分子持续移动。22S动力蛋白似乎能感知ATP浓度的细微变化,从持续模式切换到非持续模式。这些观察结果表明,动力蛋白的持续运动能力、最大力和步长与驱动蛋白相似,但ATP浓度依赖性、力-速度关系和向后位移与驱动蛋白明显不同。