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The mechanism regulating the activity of dynein, an oscillatory motor protein.

调节动力蛋白（一种振荡运动蛋白）活性的机制。

基本信息

批准号：
11440243
负责人：
SHINGYOJI Chikako
金额：
$ 8.26万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11440243/
关键词：
dynein sliding movement force oscillation central pair flagella sea urchin sperm 力発生 1分子計測

项目摘要

A prominent feature of flagellar motility is an oscillation. Flagellar oscillatory movement is based on the activity of dynein, which is one of the microtubule motor proteins. In this study I aimed to elucidate the mechanism regulating the dynein activity in the flagellar axonemes. I used sea urchin sperm flagella, which were demembranated, fragmented and then treated with elastase. A flash photolysis of caged ATP induced sliding of the elastase-treated axonemes into two groups of doublets, one of them contained the central pair. I investigated an effect of bending a part of these doublet groups and found that the bending induced two characteristic changes. One was an increase in the velocity of sliding between the two groups of the doublets. Another change was a sliding pattern, that is, the sliding occurred between any two of the nine doublets. These results indicate that the microtubule bending is involved in the regulation of dynein activity. In the second experiment, an effect of the central pair on dynein activity was studied by using singlet microtubules, which interacted with exposed dynein arms on doublets of the doublet groups. I found that the presence of the central pair inhibited the sliding activity of dynein at high calcium conditions. In the third experiment, effects of mechanical perturbation on the oscillatory force of a single dynein arm was measured and the oscillatory force was reveled to be changed by the mechanical changes applied to the doublet or the microtubules.

鞭毛运动的一个显著特征是振荡。鞭毛的振荡运动是基于动力蛋白的活性，动力蛋白是微管马达蛋白之一。本研究旨在阐明鞭毛轴丝中动力蛋白活性的调控机制。我用的是海胆精子鞭毛，它被去膜，破碎，然后用弹性蛋白酶处理。闪光光解笼ATP诱导弹性蛋白酶处理的轴丝滑成两组的双峰，其中一个包含中央对。我研究了弯曲这些双重组的一部分的效果，发现弯曲引起两个特征的变化。一个是两组双峰之间的滑动速度增加。另一个变化是滑动模式，即滑动发生在九个双峰中的任何两个之间。这些结果表明微管弯曲参与了动力蛋白活性的调节。在第二个实验中，通过使用单线态微管研究了中心对对动力蛋白活性的影响，所述单线态微管与暴露的动力蛋白臂在双峰组的双峰上相互作用。我发现，在高钙条件下，中心对的存在抑制了动力蛋白的滑动活性。在第三个实验中，我们测量了机械扰动对单个动力蛋白臂的振荡力的影响，揭示了施加到双联体或微管上的机械变化会改变振荡力。