Single-molecule measurement of a biological rotary motor, ATP synthase, driven by proton motive force

• Project/Area Number: 18770134
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Biophysics
• Research Institution: Osaka University
• Principal Investigator: IINO Ryota Osaka University, 産業科学研究所, 助教 (70403003)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,840,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥240,000); Fiscal Year 2008: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2007: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2006: ¥2,000,000 (Direct Cost: ¥2,000,000)
• Keywords: 1分子計測 / 操作 / 生体分子モーター / ATP合成酵素 / 1分子計測・操作 / 分子モーター

Research Abstract

1. 大腸菌由来のF_0F_1-ATP合成酵素(EF_0F_1)のATP合成活性を多分子計測により徹底的に調べ、回転子を構成するεサブユニットがATP加水分解時のみでなくATP合成時においても、基質の結合と生成物の解離を抑制することを明らかにし、阻害の新規モデルを提案した。
2. 基板支持人工脂質二重膜に再構成したEF_0F_1の回転(ATP加水分解駆動)を直接可視化することに成功した。さらにF_0モーターのプロトン輸送が低下した変異体を作製し、プロトン輸送律速の36°ステップを直接観察することに世界で初めて成功した。

Research Products

• [Journal Article] Mechanism of inhibition by C-terminal α-helices of the ε subunit of Escherichia coli FoF1-ATP synthase (2009)
  • Author(s): Iino R, Hasegawa R, Tabata KV, Noji H.
  • Journal Title: J. Biol. Chem. (published)
  • Peer Reviewed
• [Journal Article] Correlation between the conformational states of F1-ATPase as determined from its crystal structure and single-molecule rotation (2008)
  • Author(s): Okuno D, Fujisawa R, Iino R, Hirono-Hara Y, Imamura H, Noji H.
  • Journal Title: Proc. Natl. Acad. Soc. U.S.A. 105 Pages: 20722-20727
  • Peer Reviewed
• [Journal Article] Highly-sensitive Restriction Enzyme Assay and Analysis : A Review (2008)
  • Author(s): Lam L, Iino R, Tabata KV, Noji H
  • Journal Title: Analytical and Bioanalytical Chemistry 39 Pages: 2423-2432
  • Peer Reviewed
• [Journal Article] Temperature- sensitive reaction intermediate of F_1-ATPase (2008)
  • Author(s): Watanabe R, Iino R, Shimabukuro K, Yoshida M, Noji H.
  • Journal Title: EMBO Rep. 9 Pages: 84-90
  • Peer Reviewed
• [Journal Article] Correlation between the conformational states of F_1-ATPase as determined from its crystal structure and single-molecule rotation. (2008)
  • Author(s): Okuno D, Fujisawa R, Iino R, Hirono-Hara Y, Imamura H, *Noji H
  • Journal Title: Proc. Natl. Acad. Soc. U.S.A. 105 Pages: 20722-20727
  • Peer Reviewed
• [Journal Article] Temperature-sensitive reaction intermediate of F_1-ATPase (2008)
  • Author(s): Watanabe R, lino R, Shimabukuro K, Yoshida M, *Noji H.
  • Journal Title: EMBO Report 39 Pages: 84-90
  • Peer Reviewed
• [Journal Article] Highly-sensitive Restriction Enzyme Assay and Analysis : A Review (2008)
  • Author(s): *Lam L, lino R, Tabata KV, and *Noji H.
  • Journal Title: Analytical and Bioanalytical Chemistry 39 Pages: 2423-2432
  • Peer Reviewed
• [Journal Article] Temperature-sensitive reaction intermediate of F_1-ATPase (2008)
  • Author(s): Rikiya Watanabe
  • Journal Title: EMBO report 9 Pages: 84-90
  • Peer Reviewed
• [Journal Article] 超微小反応チャンバーを用いた高感度バイオアッセイ (2007)
  • Author(s): 飯野亮太, Liza Lam, 野地博行
  • Journal Title: 未来材料 7 Pages: 14-20
  • NAID: 40015642974
• [Journal Article] GPI-anchored receptor clusters transiently recruit Lyn and G_α for temporary cluster immobilization and Lyn activation: single-molecule tracking study 1 (2007)
  • Author(s): Suzuki KGN, Fujiwara TK, Sanematsu F, Iino R, Edidin M, Kusumi A.
  • Journal Title: J. Cell Biol. 177 Pages: 717-730
  • Peer Reviewed
• [Journal Article] 超微小反応チャンバーを用いた高感度バイオアッセイ (2007)
  • Author(s): 飯野亮太
  • Journal Title: 未来材料 7 Pages: 14-20
  • NAID: 40015642974
• [Journal Article] 超微小反応チャンバーで生体回転分子モーターの作動機構を探る (2006)
  • Author(s): 飯野亮太, 野地博行
  • Journal Title: 未来材料 6 Pages: 32-38
  • NAID: 40007316605
• [Journal Article] F_1-ATPase : A highly coupled reversible rotary motor (2006)
  • Author(s): Iino R, Noji H.
  • Journal Title: Biochem. Soc. Trans. 34 Pages: 993-996
• [Journal Article] F_1-ATPase : A highly coupled reversible rotary motor (2006)
  • Author(s): Ryota Iino, Hiroyuki Noji
  • Journal Title: Biochemical Society Transaction 34 Pages: 993-996
• [Journal Article] 超微小反応チャンバーで生体回転分子モーターの作動機構を探る (2006)
  • Author(s): 飯野亮太, 野地博行
  • Journal Title: 未来材料 6巻5号 Pages: 32-38
  • NAID: 40007316605
• [Presentation] Single-molecule Imaging of ATP-driven Stepping Rotation of F_0F_1-ATP Synthase Reconstituted into Supported Membrane (2009)
  • Author(s): Iino R, Tham K. C, Tabata K. V., Ueno H., Noji H.
  • Organizer: Biophysical Society 53rd Annual Meeting
  • Place of Presentation: Boston USA
  • Year and Date: 2009-03-01
• [Presentation] Single-molecule Imaging of ATP-driven Stepping Rotation of F_0F_1-ATP Synthase Reconstituted into Supported Membrane (2009)
  • Author(s): *Ryota lino, Khek-Chian Tham, Kazuhito V. Tabata, Hiroshi Ueno, *Hiroyuki Noji
  • Organizer: Biophysical Society 53rd Annual Meeting
  • Place of Presentation: Boston. USA
  • Year and Date: 2009-03-01
• [Presentation] How is the Temperature Sensitive (TS) Reaction of F_1-ATPase Coupled with its Rotation? (2009)
  • Author(s): Sawako Enoki, Ryota Iino, Rikiya Watanabe, *Hiroyuki Noji
  • Organizer: Biophysical Society 53rd Annual Meeting
  • Place of Presentation: Boston. USA
  • Year and Date: 2009-03-01
• [Presentation] ATP-driven rotation of F_0F_1-ATP synthase rate-limited by F_0 sector (2008)
  • Author(s): Iino R, Tham K. C, Tabata K. V., Ueno H., Noji H.
  • Organizer: 日本生物物理学会第46回年会
  • Place of Presentation: 福岡
  • Year and Date: 2008-12-05
• [Presentation] 超高感度バイオセンシング技術 (2008)
  • Author(s): 飯野亮太
  • Organizer: 産業科学研究協会 平成20年度第3回産研テクノサロン
  • Place of Presentation: 大阪
  • Year and Date: 2008-10-30
• [Presentation] 超高感度バイオセンシング技術 (2008)
  • Author(s): *飯野亮太
  • Organizer: 産業科学研究協会平成20年度第3回産研テクノサロン
  • Place of Presentation: 大阪
  • Year and Date: 2008-10-30
• [Presentation] Single-molecule study on the rotation of F_0F_1-ATPase (2008)
  • Author(s): Iino R.
  • Organizer: 5th NSF-MEXT Young Researcher Exchange Program
  • Place of Presentation: Osaka. Japan
  • Year and Date: 2008-10-12
• [Presentation] Single-molecule studies on the rotation of F_0F_1-ATPase (2008)
  • Author(s): Iino R.
  • Organizer: 4th HANDAI Nanoscience and Nanotechnology Symposium
  • Place of Presentation: Osaka. Japan
  • Year and Date: 2008-09-30
• [Presentation] Correlation among mechanical steps in rotation, chemical reaction, and crystal structure of F_1-ATPase unraveled by single-molecule studies (2008)
  • Author(s): Iino R.
  • Organizer: The Fourth Workshop of the UK-Japan Bionanotechnology Collaboration
  • Place of Presentation: Kobe. Japan
  • Year and Date: 2008-09-18
• [Presentation] ATP-driven rotation of F_0F_1-ATP synthase reconstituted into supported membrane (2008)
  • Author(s): Iino R. Tabata K., Ueno H., Noji H.
  • Organizer: 52nd Annual Meeting of Biophysical Society
  • Place of Presentation: CA, USA
  • Year and Date: 2008-02-05
• [Presentation] ATP-driven rotation of F_0F_1-ATP synthase reconstituted into supported membrane (2008)
  • Author(s): Ryota Iino
  • Organizer: 52nd Annual Meeting of Biophysical Society, USA
  • Place of Presentation: 米国
  • Year and Date: 2008-02-05
• [Presentation] Mechanochemical coupling of F_1-ATPase and intracellular ATP imaging (2008)
  • Author(s): Ryota Iino, Rikiya Watanabe, Daichi Okuno, *Hiromi Imamura, *Hiroyuki Noji
  • Organizer: 15th European Bioenergetics Conference (EBEC2008)
  • Place of Presentation: Dublin. Ireland
• [Presentation] 基板支持人工脂質二重膜に再構成したF_0F_1-ATP合成酵素のATP駆動による回転 (2007)
  • Author(s): 飯野亮太, 田端和仁, 上野博史, 野地博行
  • Organizer: 日本生物物理学会第45回年会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-23
• [Presentation] 基板支持人工脂質二重膜に再構成したF_0F_1-ATP合成酵素のATP駆動による回転 (2007)
  • Author(s): 飯野亮太
  • Organizer: 日本生物物理学会第45同年会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-23
• [Presentation] Highly sensitive measurement of biological reaction in femtoliter chamber array (2007)
  • Author(s): Iino R.
  • Organizer: SANKEN WORKSHOP on Nano-Bioscience at Berkeley
  • Place of Presentation: Berkley. CA. USA
  • Year and Date: 2007-03-22
• [Presentation] Femtoliter reaction chamber array for highly sensitive measurement of biological reaction (2006)
  • Author(s): Noji H, Iino R.
  • Organizer: UT Symposium on NanoBio Integration NANOBIO-TOKYO 2006
  • Place of Presentation: Tokyo Japan
  • Year and Date: 2006-12-05
• [Presentation] Micron-sized reaction chamber array for highly sensitive detection of biological reaction (2006)
  • Author(s): Iino R.
  • Organizer: SANKEN International Symposium on Nanoscience and Nanotechnology
  • Place of Presentation: Osaka Japan
  • Year and Date: 2006-09-19
• [Book] 生体膜超分子モーター-細胞の回転エネルギー変換装置 : ATP合成酵素.「超分子サイエンス」, 国武豊喜監修.第4章第1節第1項 (2009)
  • Author(s): 飯野亮太, 田端和仁, 野地博行
  • Publisher: NTS
• [Book] 単一分子計測.「マイクロ・ナノ化学チップと医療・環境・バイオ分析」, 北森武彦監修.第5編応用技術、第15章 (2009)
  • Author(s): 飯野亮太, 野地博行
  • Publisher: 技術教育出版
• [Book] 「マイクロ・ナノ化学チップと医療・環境・バイオ分析」第5編応用技術、第15章単-分子計測 (2009)
  • Author(s): *飯野亮太,*野地博行
  • Total Pages: 12
  • Publisher: 技術教育出版
• [Book] Two rotary motors in ATP-synthase. In "Single Molecule Dynamics in Life Science" edited by Yanagida T and Ishii Y. WILEY-VCH (Germany) (2008)
  • Author(s): Iino R, Noji H.
• [Book] 2本脚の生体分子モーターはどう歩く?MyosinVの歩きのメカニズム.「最新分子マシン.ナノで働く"高度な機械"を目指して」 (2008)
  • Author(s): 飯野亮太
  • Publisher: 化学同人
• [Book] "Single Molecule Dynamics in Life Science" Chapter 10. Two rotary motors in ATP-synthase (2008)
  • Author(s): *Iino R and *Noji H.
  • Total Pages: 19
  • Publisher: WILEY-VCH (Germany)
• [Book] 「最新分子マシン.ナノで働く"高度な機械"を目指して」第4章5節.2本脚の生体分子モーターはどう歩く? MyosinVの歩きのメカニズム (2008)
  • Author(s): *飯野亮太
  • Total Pages: 2
  • Publisher: 化学同人
• [Book] 基礎ナノバイオテクノロジー
  • Author(s): 飯野亮太, 野地博行ら
  • Publisher: 培風館(印刷中)
• [Remarks] 飯野亮太 電気学会 2007年度研究会 優秀論文発表賞(センサ・マイクロマシン部門)
• [Patent(Industrial Property Rights)] Polarized total internal reflection illumination optical system by rotary annulus light (2009)
  • Inventor(s): Masasuke Yoshida, Toshiharu Suzuki, Megumu Shio, Ryota Iino
  • Industrial Property Rights Holder: Japan Science and Technology Agency
  • Filing Date: 2009-02-03
  • Overseas
• [Patent(Industrial Property Rights)] 回転式輪帯全反射偏光照明光学系 (2009)
  • Inventor(s): 吉田賢右, 鈴木俊治, 塩育, 飯野亮太
  • Industrial Property Rights Holder: 独立行政法人科学技術振興機構
  • Filing Date: 2009-01-16
• [Patent(Industrial Property Rights)] 回転式輪帯全反射偏光照明光学系 (2009)
  • Inventor(s): 吉田賢右、鈴木俊治、塩育、*飯野亮太
  • Industrial Property Rights Holder: 科学技術振興機構
  • Acquisition Date: 2009-01-16
• [Patent(Industrial Property Rights)] アデノシン三リン酸測定のための蛍光標識タンパク質 (2007)
  • Inventor(s): 今村博臣, 山田康之, 飯野亮太, 野地博行
  • Industrial Property Rights Holder: 財団法人新産業創造研究機構, 国立大学法人大阪大学, 学校法人立教学院
  • Industrial Property Number: 2007-008256
  • Filing Date: 2007-03-27
• [Patent(Industrial Property Rights)] 細胞検体の異物輩出活性検出法、およびその利用 (2006)
  • Inventor(s): 飯野亮太, 西野邦彦, 仲田昌義, 榊原昇一, 山口明人, 野地博行
  • Industrial Property Rights Holder: 国立大学法人大阪大学
  • Industrial Property Number: 2006-294558
  • Filing Date: 2006-10-30