Single molecule analysis of transport proteins

• Project/Area Number: 17H03660
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biophysics
• Research Institution: Institute of Physical and Chemical Research (2019); The University of Tokyo (2017-2018)
• Principal Investigator: Rikiya Watanabe 国立研究開発法人理化学研究所, 開拓研究本部, 主任研究員 (30540108)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2018: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); 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 Final Research Achievements

In this study, we succeeded to measure the conformational change and ion transport of FoF1 ATP synthase by using a novel artificial cell-membrane microsystem. In addition, we expanded the versatility of this platform to other transport proteins.

Academic Significance and Societal Importance of the Research Achievements

トランスポーターは、構造変化と共役して標的分子を輸送する膜タンパク質であり、細胞内の恒常性の維持に深く関わっている。近年、トランスポーターに関連する様々な疾患が同定され、トランスポーターの作動機構を詳細に解明することは、学術・産業の両面において急務とされている。本研究成果はトランスポーターの作動機序を理解する上で、重要な技術基盤の確立の第一歩であり、今後のトランスポーターの研究の発展に寄与することが期待される。

Research Products

• [Journal Article] Microsystem for the single molecule analysis of membrane transport proteins. (2020)
  • Author(s): Rikiya Watanabe
  • Journal Title: Biochim Biophys Acta Gen Subj Volume: 19 Issue: 2 Pages: 30072-30078
  • DOI: 10.1016/j.bbagen.2019.03.016
  • Peer Reviewed
• [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] Microsystems for single molecule analysis of membrane proteins (2019)
  • Author(s): Rikiya Watanabe
  • Organizer: 23rd International Conference on Miniaturized Systems for Chemistry and Life Sciences (microTAS2019)
  • Int'l Joint Research / Invited
• [Presentation] Microsystems for single molecule analysis of membrane proteins (2019)
  • Author(s): Rikiya Watanabe
  • Organizer: 32nd International Microprocesses and Nanotechnology Conference (MNC2019)
  • Int'l Joint Research / Invited
• [Presentation] Microsystems for single molecule analysis of membrane proteins (2019)
  • Author(s): Rikiya Watanabe
  • Organizer: 17th Chinese Biophysics Congress “China-Japan Joint Minisymposium on Single-Molecule Biophysics
  • Int'l Joint Research / Invited
• [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