Lombillo V A, Stewart R J, McIntosh J R
Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder 80309.
Nature. 1995 Jan 12;373(6510):161-4. doi: 10.1038/373161a0.
Dynamic changes in microtubule (MT) length have long been thought to contribute to intracellular motility. Both the polymerization and depolymerization of tubulin have been shown to do work in vitro, but the biochemical complexity of objects moved, such as chromosomes, has complicated the identification of proteins that couple MT dynamics with motility. Work with MTs grown from and tethered to pellicles of lysed Tetrahymena has shown that disassembly-dependent movement of chromosomes in vitro can be inhibited with antibodies against the motor domain of kinesin. To study proteins that can function in disassembly-dependent motion, we have refined this motility assay, replacing chromosomes with protein-coated latex microspheres. We report here the ability of several enzymes, including kinesin, to support in vitro motility of latex microspheres on disassembling MTs (Fig. 1a). The polarity of kinesin's motor activity can be reversed by MT disassembly and interactions between a motor and a MT end can either slow or speed the rate of tubulin depolymerization.
长期以来,人们一直认为微管(MT)长度的动态变化有助于细胞内运动。已证明微管蛋白的聚合和解聚在体外都能做功,但诸如染色体等被移动物体的生化复杂性,使得鉴定将MT动态变化与运动联系起来的蛋白质变得复杂。对从裂解的四膜虫的薄膜生长并与之相连的MT的研究表明,体外染色体的依赖于解聚的运动可以被针对驱动蛋白运动结构域的抗体所抑制。为了研究能在依赖于解聚的运动中发挥作用的蛋白质,我们改进了这种运动分析方法,用蛋白包被的乳胶微球取代了染色体。我们在此报告了几种酶,包括驱动蛋白,支持乳胶微球在解聚的MT上进行体外运动的能力(图1a)。驱动蛋白的运动活性极性可通过MT解聚而逆转,并且一个驱动蛋白与一个MT末端之间的相互作用既可以减慢也可以加快微管蛋白解聚的速率。