Structural basis of ATP hydrolysis mechanism of V1-ATPase motor

V1-ATP酶马达ATP水解机制的结构基础

• 批准号: 23370047
• 负责人: MURATA Takeshi
• 金额: $ 9.57万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23370047/
• 关键词: 分子モーター; 酵素反応; V-ATPase; X線結晶構造解析; ATP

Vacuolar ATPases (V-ATPases) function as proton pumps in various membrane systems within the cells, which are involved in a number of processes such as bone resorption and cancer metastasis. The hydrophilic V1 portion is known as a rotary motor, in which a central axis DF complex rotates inside a hexagonally arranged catalytic A3B3 complex using ATP hydrolysis energy, although the molecular mechanism is not well defined due to a lack of high-resolution structural information.We have established the in vitro expression, purification, and reconstitution of Enterococcus hirae V1-ATPase from the A3B3 and DF complexes. In this study, we solved the crystal structures of AMP-PNP-bound and Pi-bound V1-ATPase, and also characterized the several site-directed mutants. On the basis of these findings, we proposed the molecular mechanism of the rotary motor.

极性ATP酶（V-ATP酶）在细胞内的各种膜系统中起质子泵的作用，其参与许多过程，如骨吸收和癌症转移。亲水性V1部分被称为旋转马达，其中中心轴DF复合物利用ATP水解能量在六边形排列的催化A3 B3复合物内旋转，尽管由于缺乏高分辨率的结构信息，分子机制还没有很好地定义。在本研究中，我们解决了AMP-PNP结合和Pi结合的V1-ATP酶的晶体结构，并表征了几个定点突变体。在此基础上，提出了旋转马达的分子机理。

项目成果

Crystal structure of the central axis DF complex of the prokaryotic V-ATPase

• DOI: 10.1073/pnas.1108810108
• 发表时间: 2011-12-13
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Saijo, Shinya;Arai, Satoshi;Murata, Takeshi
• 通讯作者: Murata, Takeshi

Rotation mechanism of Enterococcus hirae V1-ATPase based on asymmetric crystal structures

• DOI: 10.1038/nature11778
• 发表时间: 2013-01-31
• 期刊: NATURE
• 影响因子: 64.8
• 作者: Arai, Satoshi;Saijo, Shinya;Murata, Takeshi
• 通讯作者: Murata, Takeshi

V-ATPaseの分子メカニズム, 膜タンパク質構造研究

V-ATPase分子机制、膜蛋白结构研究

• 发表时间: 2013
• 作者: 水谷健二;山本三沙岐;村田武士
• 通讯作者: 村田武士

Crystallization of membrane proteins using functional antibody fragments

使用功能性抗体片段结晶膜蛋白

• 发表时间: 2012
• 作者: 武島弘彦;ら;Takeshi Murata
• 通讯作者: Takeshi Murata

Crystal structure of V1-ATPase from Enterococcus hirae

海拉肠球菌 V1-ATP 酶的晶体结构

• 发表时间: 2011
• 作者: Tokuda;N.;Igarashi;K.;Shimamura;T.;Yurugi-Kobayashi;T.;Shiroishi;M.;Ito;K.;Sugawara;T.;Asada;H.;Murata;T.;Nomura;N.;Iwata;S. and Kobayashi;T;Takeshi Murata
• 通讯作者: Takeshi Murata