权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Energy Transduction in Sodium Driven-flagagellar Motor

钠驱动鞭毛运动中的能量转换

基本信息

批准号：
09480174
负责人：
HOMMA Michio
金额：
$ 8.13万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09480174/
关键词：
ion channel energy transduction Vibrio flagella motor Sodium motive force

项目摘要

Four motor proteins, MotX, MotY, PomA and PomB have been identified in the Na^+-type flagella of Vibrio spp. They are integral membrane proteins and essential components for the rotation of the Na^+-driven polar flagellar motor of Vibrio alginolyticus. To investigate PomA function, we carried out random mutagenesis of the pomA gene by using hydroxylamine. As the results, it is suggested that PomA and MotA of H^+ motor have very similar structure and roles. So the basic mechanism for torque generation will be similar in the proton and sodium motor. The complete motX gene was cloned from Vibrio alginolyticus and was shown to complement three mot mutations, motX94, motX115 and motX119, as well as a motX mutant of V.parahaemolyticus. The study of the flagellar motor has been intensively done in the H^+-motor and there are five core proteins (MotA, MotB, FliG, FliM and FliN) involved in the motor function. In the rotor part of the motor, three soluble proteins, FliG, FliM and FliN, are the components for force-generation, flagellar assembly and controlling the direction of motor rotation. Among them, FliG has a direct role in force generation with the cytoplaslmic region of MotA.We have clnoed the fliG gene of Na^+-driven polar flagellar motor and the product could be purified using the His tag. To verify whether the motor proteins form a complex, immunoprecipitation was carried out. PomB was coprecipitated with PomA by anti-PomA antibody, and anti-PomB antibody coprecipitated with PomA as well. Those directly demonstrated that PomA and PomB functionally interact with each other.

在弧菌的Na~+型鞭毛中已鉴定出MotX、Moty、Poma和Pomb四种运动蛋白。它们是完整的膜蛋白，也是溶藻弧菌由Na~+驱动的极鞭毛马达旋转的重要组成部分。为了研究POMA的功能，我们用羟胺对POMA基因进行了随机突变。结果表明，H~+马达的POMA和MOTA具有非常相似的结构和作用。因此，在质子和钠发动机中产生扭矩的基本机制将是相似的。从溶藻弧菌中克隆了完整的motX基因，该基因与motX94、motX115和motX119三个mot突变株以及副溶血弧菌的一个motX突变株具有互补作用。对鞭毛马达的研究主要集中在H^+-马达中，有五种核心蛋白(MOTA、MOTB、FliG、FLiM和FLIN)参与运动功能。在电机的转子部分，三种可溶性蛋白质FliG、FliM和Flin是力产生、鞭毛组装和控制电机旋转方向的组件。其中，FliG与Mota的细胞质区域具有直接的作用力作用。我们克隆了Na~+驱动的极鞭毛马达的FliG基因，并利用His标签对其进行了纯化。为了验证马达蛋白是否形成复合体，进行了免疫沉淀。Pomb与POMA通过抗POMA抗体共沉淀，抗Pomb抗体也与POMA共沉淀。这直接证明了庞玛和庞贝在功能上是相互作用的。