Drosophila kinesin: characterization of microtubule motility and ATPase.
Drosophila kinesin: characterization of microtubule motility and ATPase.
复制标题
果蝇驱动蛋白：微管运动和 ATP 酶的表征。
DOI：
10.1073/pnas.85.4.1109
复制
发表时间：
1988
影响因子：
11.1
通讯作者：
McIntosh,JR
中科院分区：
文献类型：
--
作者：
Saxton,WM;Porter,ME;Cohn,SA;Scholey,JM;Raff,EC;McIntosh,JR
通过将微管与不可水解的 ATP 类似物一起孵育，并对 ATP 从微管中释放的蛋白质进行凝胶过滤，从果蝇胚胎中分离出了驱动蛋白的制剂，驱动蛋白是一种基于微管的力产生蛋白。这些制剂在体外诱导 MgATP 依赖性微管滑动，MgATP 的 Km 为 44 microM，滑动的 Vmax 为 0.9 微米/秒。在运动测定中具有活性的果蝇蛋白质样品在溶液中的平均 ATP 酶活性为每毫克 17 纳摩尔/分钟，在微管存在的情况下，该活性增加到每毫克平均 106 纳摩尔/分钟。与这些活性共纯化的主要多肽显示出115 kDa 和58 kDa 的相对分子质量。针对115-kDa多肽产生的抗血清还识别鱿鱼驱动蛋白制剂的110-kDa成分和海胆驱动蛋白制剂的130-kDa成分。
Preparations of kinesin, a microtubule-based force-producing protein, have been isolated from Drosophila melanogaster embryos by incubation of microtubules with a nonhydrolyzable ATP analogue and gel filtration of proteins released from the microtubules by ATP. These preparations induced MgATP-dependent microtubule gliding in vitro with a Km for MgATP of 44 microM and a Vmax for gliding of 0.9 micron/sec. Samples of Drosophila proteins that were active in motility assays possessed an average ATPase activity in solution of 17 nmol/min per mg that increased to an average of 106 nmol/min per mg in the presence of microtubules. The major polypeptides that copurified with these activities showed relative molecular masses of 115 kDa and 58 kDa. An antiserum raised against the 115-kDa polypeptide also recognized the 110-kDa component of squid kinesin preparations and the 130-kDa component of sea urchin kinesin preparations.