擬似細胞チップによる膜タンパク質の動的構造機能計測技術の開発

• Project Area: Novel measurement techniques for visualizing 'live' protein molecules at work
• Project/Area Number: 17H05871
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: The University of Tokyo
• Principal Investigator: 渡邉 力也 東京大学, 大学院工学系研究科(工学部), 客員研究員 (30540108)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥13,000,000 (Direct Cost: ¥10,000,000、Indirect Cost: ¥3,000,000); Fiscal Year 2018: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2017: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 1分子生物物理 / 膜タンパク質 / トランスポーター / MEMS / bioMEMS

Outline of Annual Research Achievements

本研究では、疑似細胞アレイチップを作成し、細胞膜機能を人工的に再構築するとともに、膜タンパク質の構造や機能の高感度計測基盤の開発を行った。具体的には、細胞膜画分をマイクロチップ上の生体膜と融合させることで、膜タンパク質を機能を保持した状態で再構成することが可能となり、特に膜輸送体に関して様々な機能を定量的に計測する基盤技術を確立することに成功した。また、F型ATP合成酵素などの構造変化の可視化技術の開発にも成功した。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] High-throughput single-molecule bioassay using micro-reactor arrays with a concentration gradient of target molecules (2018)
  • Author(s): Watanabe Rikiya、Komatsu Toru、Sakamoto Shingo、Urano Yasuteru、Noji Hiroyuki
  • Journal Title: Lab on a Chip Volume: 18 Issue: 18 Pages: 2849-2853
  • DOI: 10.1039/c8lc00535d
  • Peer Reviewed / Open Access
• [Journal Article] Single-molecule analysis of phospholipid scrambling by TMEM16F. (2018)
  • Author(s): Watanabe, R., Sakuragi, T., Noji,H. and Nagata, S.
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 115 Issue: 12 Pages: 3066-3071
  • DOI: 10.1073/pnas.1717956115
  • Peer Reviewed / Open Access
• [Journal Article] Essential Role of the ε Subunit for Reversible Chemo-Mechanical Coupling in F 1 -ATPase (2018)
  • Author(s): Watanabe Rikiya、Genda Makoto、Kato-Yamada Yasuyuki、Noji Hiroyuki
  • Journal Title: Biophysical Journal Volume: 114 Issue: 1 Pages: 178-187
  • DOI: 10.1016/j.bpj.2017.11.004
  • Peer Reviewed
• [Journal Article] Single-Molecule Analysis of Membrane Transporter Activity by Means of a Microsystem (2017)
  • Author(s): Watanabe Rikiya、Soga Naoki、Ohdate Shin-ya、Noji Hiroyuki
  • Journal Title: Methods in Molecular Biology Bacterial Multidrug Exporters Volume: 1700 Pages: 321-330
  • DOI: 10.1007/978-1-4939-7454-2_17
  • ISBN: 9781493974528, 9781493974542
  • Peer Reviewed / Open Access
• [Journal Article] Effects of non-equilibrium angle fluctuation on F1-ATPase kinetics induced by temperature increase (2017)
  • Author(s): Yuji Tamiya, Rikiya Watanabe, Hiroyuki Noji, Chun-Biu Li, Tamiki Komatsuzaki
  • Journal Title: Physical Chemistry Chemical Physics Volume: 3(20) Issue: 3 Pages: 1872-1880
  • DOI: 10.1039/c7cp06256g
  • NAID: 120006549169
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Artificial cell-membrane microsystems for highly sensitive analysis of membrane proteins (2018)
  • Author(s): Rikiya Watanabe
  • Organizer: PITTCON 2018
  • Int'l Joint Research / Invited
• [Presentation] 生体膜マイクロチップを利用した膜タンパク質の機能可視化 (2018)
  • Author(s): 渡邉力也
  • Organizer: 日本細菌学会年会
  • Invited
• [Presentation] Single molecule analysis of membrane transport proteins using artificial cell-membrane microsystems (2017)
  • Author(s): 渡邉力也
  • Organizer: 日本生物物理学会年会
  • Invited
• [Presentation] 生体膜チップを利用した 膜タンパク質の1分子活性計測 (2017)
  • Author(s): 渡邉力也
  • Organizer: Conbio 2017
  • Invited