Molecular mechanism of the structural formation and functional regulation of the V1 rotary motor

• Project/Area Number: 26291009
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Chiba University
• Principal Investigator: Murata Takeshi 千葉大学, 大学院理学研究科, 教授 (80415322)
• Co-Investigator(Renkei-kenkyūsha): MIZUTANI Kenji 横浜市立大学, 生命医科学研究科, 助教 (10525570) ; YAMATO Ichiro 東京理科大学, 基礎工学部, 教授 (70111458)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2015: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000); Fiscal Year 2014: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000)
• Keywords: V-ATPase / 回転分子モーター / X線結晶構造解析 / 分子モーター / 構造解析

Outline of Final Research Achievements

V-ATPases function as ATP-dependent ion pumps, the hydrophilic V1 portion is known as rotary motor in which a central axis DF complex rotates inside a hexagonally arranged catalytic A3B3 complex using ATP hydrolysis energy. We previously succeeded in obtaining the crystal structures of the A3B3 complex which is a three-fold assembly of the identical A1B1 pair, but showed asymmetrical hexamer ring structure. In this study, we elucidated the crystal structures and biochemical properties of the A-subunit, B-subunit, A1B1 complex, and A3B3 mutant. Based on these and previous findings, we propose a molecular mechanism model of the structural formation and functional regulation of the V1 rotary motor.

Research Products

• [Journal Article] Crystal structures of the ATP-binding and ADP-release dwells of the V1 rotary motor. (2016)
  • Author(s): Kano Suzuki, Kenji Mizutani, Shintaro Maruyama, Kazumi Shimono, Fabiana L. Imai, Eiro Muneyuki, Yoshimi Kakinuma, Yoshiko Ishizuka-Katsura, Mikako Shirouzu, Shigeyuki Yokoyama, Ichiro Yamato, and Takeshi Murata
  • Journal Title: Nature Communications Volume: 7 Issue: 1 Pages: 13235-13235
  • DOI: 10.1038/ncomms13235
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Operating principles of rotary molecular motors: differences between F₁ and V₁ motors (2016)
  • Author(s): Ichiro Yamato, Yoshimi Kakinuma, Takeshi Murata
  • Journal Title: Biophysics and Physicobiology Volume: 13 Issue: 0 Pages: 37-44
  • DOI: 10.2142/biophysico.13.0_37
  • NAID: 130005129245
  • ISSN: 2189-4779
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Rotational mechanism of Enterococcus hirae V1-ATPase by crystal-structure and single-molecule analysis (2015)
  • Author(s): Iino R, Minagawa Y, Ueno H, Suzuki K, and Murata T
  • Journal Title: Current Opinion in Structural Biology Volume: 31 Pages: 49-56
  • DOI: 10.1016/j.sbi.2015.02.013
  • Peer Reviewed
• [Journal Article] Infrared spectroscopic studies on the V-ATPase (2015)
  • Author(s): H. Kandori*, Y. Furutani, T. Murata
  • Journal Title: Biochim. Biophys Acta Volume: 1847 Issue: 1 Pages: 134-141
  • DOI: 10.1016/j.bbabio.2014.07.020
  • Peer Reviewed
• [Journal Article] Torque generation of Enterococcus hirae V-ATPase (2014)
  • Author(s): Ueno H, Minagawa Y, Hara M, Rahman S, Yamato I, Muneyuki E, Murata T, and Iino R
  • Journal Title: The Journal of Biological Chemistry Volume: 289 Issue: 45 Pages: 31212-31223
  • DOI: 10.1074/jbc.m114.598177
  • Peer Reviewed / Open Access
• [Journal Article] Molecular structure and rotary dynamics of Enterococcus hirae V1-ATPase (2014)
  • Author(s): Iino R, Minagawa Y, Ueno H, Hara M, and Murata T
  • Journal Title: IUBMB Life Volume: 66 Issue: 9 Pages: 624-630
  • DOI: 10.1002/iub.1311
  • Peer Reviewed
• [Journal Article] 膜超分子モーター（V-ATPase）の分子メカニズム (2014)
  • Author(s): 村田武士
  • Journal Title: 実験医学 Volume: 32 Pages: 1623-1626
• [Journal Article] 非対称結晶構造からみえてきたV1-ATPaseの回転メカニズム (2014)
  • Author(s): 鈴木花野、村田武士
  • Journal Title: 医学のあゆみ Volume: 249 Pages: 551-552
• [Presentation] 結晶構造からわかる回転機動分子の機能発現機構 (2016)
  • Author(s): 村田武士
  • Organizer: 日本化学会第96春季年会
  • Place of Presentation: 同志社大学
  • Year and Date: 2016-03-27
  • Invited
• [Presentation] 膜蛋白質の耐熱化と発現・精製 (2016)
  • Author(s): 村田武士
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: 福岡国際会議場（福岡県・福岡市）
  • Invited
• [Presentation] Ion transporting mechanism of V-ATPase (2015)
  • Author(s): Takeshi Murata
  • Organizer: 第53回日本生物物理学会
  • Place of Presentation: 金沢大学
  • Year and Date: 2015-09-13
  • Invited
• [Presentation] Rotational mechanism of Enterococcus hirae V1-ATPase (2015)
  • Author(s): Takeshi Murata
  • Organizer: Gordon Research Conference in Molecular and Cellular Bioenergetics
  • Place of Presentation: Proctor academy
  • Year and Date: 2015-06-22
  • Int'l Joint Research / Invited
• [Presentation] 創薬標的膜タンパク質の構造解析法について (2015)
  • Author(s): 村田武士
  • Organizer: 第５６回日本生化学会中国四国支部例会
  • Place of Presentation: 島根大学
  • Year and Date: 2015-05-29
  • Invited
• [Presentation] ヒト膜タンパク質の大量発現・精製・結晶化の技術開発 (2014)
  • Author(s): 村田 武士
  • Organizer: 平成26年度日本結晶学会年会
  • Place of Presentation: 東京大学農学部 弥生講堂・一条ホール
  • Year and Date: 2014-11-03
  • Invited
• [Presentation] Crystallization methods of membrane proteins (2014)
  • Author(s): 村田 武士
  • Organizer: 第52回日本生物物理学会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2014-09-25
  • Invited
• [Presentation] 相関構造解析によるV１モーターの回転メカニズム (2014)
  • Author(s): 村田 武士
  • Organizer: 第14回蛋白質科学会
  • Place of Presentation: ワークピア横浜／横浜産貿ホール
  • Year and Date: 2014-06-25
  • Invited
• [Book] Function and Regulation of Mammalian V-ATPase Isoforms, Regulation of Ca2+-ATPase, V-ATPases and F-ATPases, Advances in Biochemistry in Health and Disease 14 (2016)
  • Author(s): Suhaila Rahman, Ichiro Yamato, Takeshi Murata
  • Total Pages: 17
  • Publisher: Springer International Publishing