Mechanical Regulation of V type Rotary Motor

• Project/Area Number: 18370055
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: YOKOYAMA Kin Tokyo Institute of Technology, CHEMICAL RESOURCE LABORATORY, SPECIAL RESERCHER (70271377)
• Co-Investigator(Kenkyū-buntansha): NOJI Hiroyuki OSAKA UNIVERSITY, THE INSTITUTE OF SCIENCE AND INDUSTRIAL RESEARCH, PROFESSOR (00343111)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥16,420,000 (Direct Cost: ¥15,700,000、Indirect Cost: ¥720,000); Fiscal Year 2007: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2006: ¥13,300,000 (Direct Cost: ¥13,300,000)
• Keywords: V-ATPASE / ATP / ATP SYNTHESIS / ROTARY MOTOR

Research Abstract

ATP synthesis by V-ATPase from the thermophilic bacterium Thermus thermophilus driven by the acid-base transition was investigated. The rate of ATP synthesis increased in parallel with increase in proton motive force (PMF) above 110 mV, which is composed of a difference in proton concentration (ΔpH) and the electrical potential differences (ΔΨ) across membranes. The optimum rate of synthesis reached 85 s^<-1> and the H^+/ATP ratio of 4.0± 0.1 was obtained. ATP was synthesized at a considerable rate solely by ΔpH indicating ΔΨ was not absolutely required for synthesis. Consistent with the H^+/ATP ratio, cryo-electron micrograph images of two-dimensional crystals of the membrane-bound rotor ring of the V-ATPase at 7.0 A resolution showed the presence of 12 V_o-c subunits each composed of two transmembrane helices. These results indicate that symmetry mismatch between the rotor and catalytic domains is not obligatory for rotary ATPases/synthases.

Research Products

• [Journal Article] ATP hydrolysis and synthesis of a rotary motor V-ATPases from Thermus thermophilus (2008)
  • Author(s): Masahiro Nakano, Hiromi Imamura, Masashi Toei, Masatada Tamakoshi, Masasuke Yoshida, and Ken Yokoyama
  • Journal Title: The Journal of Biological Chemistry 283(In press)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Molecular mechanism of energy conservation in polysulfide respiration (2008)
  • Author(s): Jormakka, M., Yokoyama, K., Yano, T., Tamakoshi, M., Akimoto, S., Shimamura, T., Curmi, P., and Iwata, S
  • Journal Title: Nature Structure and Molecular Biology 15(In press)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Molecular mechanism of energy conservation in polysulfide respiration (2008)
  • Author(s): Jormakka M., Yokoyama, K. Yano, T., Tamakoshi, M., Akimoto, S., Shimamura, T. Curmi, P., and Iwata, S
  • Journal Title: Nat Struct Mol Biol. 15
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] ATP hydrolysis and synthesis of a rotary motor V-ATPases from Thermus thermophilus (2008)
  • Author(s): Masahiro, Nakano, Hiromi, Imamura, Masashi, Toei, Masatada, Tamakoshi, Masasuke, Yoshida, Ken, Yokovama
  • Journal Title: JBiol Chem (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dodecamer rotor ring defines H^+/ATP ratio for ATP synthesis of prokaryotic V-ATPase from Thermus thermophilus (2007)
  • Author(s): Toei, H., Gerle, C., Nakano, N., Tani, K., Gyobu, N., Tamakoshi, M., Nobuhito Sone, N., Yoshida, M., Fujiyoshi, Y., *Mitsuoka, K., and Yokoyama K
  • Journal Title: The Proceedings of the National Academy of Sciences of the United States of America 103 Pages: 20256-20261
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Dodecamer rotor ring defines H^+/ATP ratio for ATP synthesis of prokaryotic V-ATPase from Thermus thermophilus Toei (2007)
  • Author(s): H., Gerle, C., Nakano, N., Tani, K., Gyobu, N., Tamakoshi, M., Nobuhito, N., Sone, M., Yoshida, Y., Fujiyoshi, K., *Mitsuoka, K., *Yokoyama
  • Journal Title: Proc Nati Acad Sci USA. 103 Pages: 20256-20261
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dodecamer rotor ring defines H^+/ATP ratio for ATP synthesis of prokaryotic V-ATPase from Thermus thermophilus (2007)
  • Author(s): Toei, H., Gerle, C., Nakano, N., Tani, K., Gyobu, N., Tamakoshi, M., Nobuhito Sone, N., Yoshida, M., Fujiyoshi, Y., *Mitsuoka, K., and Yoko yama, K
  • Journal Title: The Proceedings of the National Academy of Sciences of the United States of America 103 Pages: 20256-20261
  • Peer Reviewed
• [Journal Article] Reconstitution in vitro of V1 complex of Thermus thermophilus V-ATPase revealed that ATP binding to the A subunit is crucial for V1 formation (2006)
  • Author(s): Imamura Hiromi, Funamoto Saeko, Yoshida Masasuke, and Yokoyama Ken
  • Journal Title: The Journal of Biological Chemistry 281 Pages: 38582-38592
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Two-dimensional Crystallization and Analysis of Projection Images of Intact Thermus thermophilus V-ATPase. (2006)
  • Author(s): Gerle, C., Tani, K., Yokoyama, K., Tamakoshi, M, Yoshida, M., Fujiyoshi, Y., Mitsuoka, K.
  • Journal Title: Journal of Structural Biology 153 Pages: 200-206
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Reconstitution in vitro of V_1 complex of Thermus thermophilus V-ATPase revealed that ATP binding to the A subunit is crucial for V_1 formation (2006)
  • Author(s): H., Imamura, S., Funamoto, M., Yoshida. K., Yokovama
  • Journal Title: JBiol Chem 281 Pages: 38582-38592
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two-dimensional Crystallization and Analysis of Projection Images of Intact Thermus thermophilus V-ATPase (2006)
  • Author(s): C., Genie K., Tani, K., Yokoyama, M. Tamakoshi, M., Yoshida, Y., Fujiyoshi, K., *Mitsuoka
  • Journal Title: J. Structural Biology 153 Pages: 200-206
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Structures of bacterial type V-ATPase/synthase (2007)
  • Author(s): Ken Yokoyama
  • Organizer: FASEB Summer Research Conference
  • Place of Presentation: Vermont Academy, Saxtons River, Vermont, USA
  • Year and Date: 2007-07-11
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] FASEB Summer Research Conference Structures of bacterial type V-ATPase/synthase (2007)
  • Author(s): Ken, Yokoyama
  • Organizer: Vermont Academy, Saxtons River, Vermont
  • Place of Presentation: USA
  • Year and Date: 2007-07-11
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] 「非侵襲.可視化技術ハンドブック-ナノ・バイオ 医療から情報システムまで-」 (2007)
  • Author(s): 横山 謙, 他
  • Total Pages: 1080
  • Publisher: エヌティーエヌ出版