Single molecule reaction analysis of P-types ATPase using electrochemical fast-scanning atomic force microscopy

使用电化学快速扫描原子力显微镜对 P 型 ATP 酶进行单分子反应分析

• 批准号: 22770148
• 负责人: YOKOKAWA Masatoshi
• 金额: $ 2.75万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22770148/
• 关键词: P 型ATPase; SERCA; Na+-Pump; 高速AFM; 一分子解析; 高速原子間力顕微鏡; P型ATPase; 筋小胞体カルシウムポンプ; ナトリウムポンプ; 一分子反応解析; 電気生理

Ion pumps transport specific cationic ion across the biomembrane against a concentration gradient through the energy obtained from ATP hydrolysis. Recent progresses in crystallographic study of sarcoplasmic reticulum Ca^<2+>-ATPase (SERCA), which is one of the best-characterized P-type ATPases, allow us to discuss the transport mechanisms involving dynamical structural changes of P-type ATPases.In this study, we constructed a novel method to analyze molecular reaction of P-type ATPase at a single molecular level and real time scale using a newly developed fast-scanning atomic force microscopy. Using the system, we succeeded to observe single molecule of SERCA and plasma membrane sodium pump act as a pumping machine by direct detection of their repeatedly up-and-down conformational changes.

离子泵通过从三磷酸腺苷(ATP)水解获得的能量，通过浓度梯度将特定的阳离子输送到生物膜上。肌浆网Ca^&lt；2+&Gt；-ATPase(SERCA)是最具代表性的P型ATPase之一，其结晶学研究的最新进展使我们能够探讨涉及P型ATPase动态结构变化的转运机制。利用该系统，我们成功地观察到单分子SERCA和质膜钠泵作为泵浦机器，通过直接检测其重复上下的构象变化。

项目成果

Electrochemical microdevices

电化学微型器件

• 作者: Yokokawa M;Itoh D;Suzuki H
• 通讯作者: Suzuki H

Protein-DNA interactions in high speed AFM: single molecule diffusion analysis of human RAD54.

• DOI: 10.1039/c1ib00039j
• 发表时间: 2011-10
• 期刊: Integrative biology : quantitative biosciences from nano to macro
• 作者: H. Sánchez;Yuki Suzuki;M. Yokokawa;K. Takeyasu;C. Wyman

Polarity orientation of individual microtubules by nanochannels

纳米通道对单个微管的极性定向

• 发表时间: 2010
• 作者: Yokokawa M;Kitamura M;Yokokawa R;Kotera H
• 通讯作者: Kotera H

Nanotechnology for highly sensitive detections

用于高灵敏度检测的纳米技术

• 作者: Yokokawa M;Itoh D;Suzuki H
• 通讯作者: Suzuki H

Biological Application of Fast-Scanning Atomic Force Microscopy

快速扫描原子力显微镜的生物学应用

• 作者: Suzuki Y;Yokokawa M;Yoshimura S;Takeyasu K
• 通讯作者: Takeyasu K