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Elucidation of the input-output characteristics of F_1-motor

Research Project

Project/Area Number 14380313
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionTokyo Institute of Technology

Principal Investigator

EIRO Muneyuki  Tokyo Institute of Technology, Chemical Resources Laboratory, research associate, 資源化学研究所, 助手 (80219865)

Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥8,500,000 (Direct Cost: ¥8,500,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2002: ¥5,900,000 (Direct Cost: ¥5,900,000)
KeywordsATP synthase / free energy / energy transduction / rotary motor / work / heat / ratchet mode / fluctuation / F_1-ATPase / モーター蛋白質
Research Abstract

Biological motor proteins convert free energy of ATP hydrolysis to mechanical work, thus playing essential roles in life. But the fundamental question how they work at different free energy of ATP hydrolysis has been unanswered. This question becomes serious when we ask how stepping motion of a single motor molecule in a medium reflects the concentrations of ATP. ADP and inorganic phosphate in their individual actions. The F_1 portion of ATP synthase, also called as F_1-ATPase, is a rotary molecular motor in which central γ subunit rotates against α_3β_3 cylinder hydrolyzing ATP. Its structure is highly stable. The rotary action of this motor is processive and generates high torque. These features are ideal to explore the relationship between input free energy and output mechanical work except that this motor protein is severely inhibited by ADP. Here we overcame this problem by introducing several mutations while keeping high enzymatic activity. With a probe of beads attached, steppin … More g rotation against viscous load was examined at a wide range of free energy by changing ADP concentration. The results showed that the work of individual step motion was not affected by the free energy of ATP hydrolysis but their frequency depended on the free energy of ATP showing a property of a molecular machine operating in an environment dominated by Brownian motion.
In addition, we have analyzed rotations of a mutant F_1 subcomplex (E190D) which hydrolyzes ATP slowly and rotations of the wild-type F_1 employing a slowly hydrolysable substrate ATPγS (adenosine-5'-(γ-thio)-triphosphate). In both cases, interim dwells at 80° position were extended. We concluded that that cleavage of ATP takes place during the interim dwell.
Furthermore, we could observe rotation under an optical microscope, while watching which of the three sites bound and released a fluorescent ATP analog. If we assume that the analog mimics authentic ATP, the following scheme emerges : (i)in the ATP-waiting state, one site, dictated by the orientation of γ, is empty whereas the other two bind a nucleotide ; (ii)ATP binding to the empty site drives 〜80° rotation of γ ; (iii)this triggers a reaction(s), hydrolysis and/or phosphate release but not ADP release, in the site that bound ATP one step ago ; (iv)completion of that reaction induces further 〜40° rotation.
Thus, we could elucidate important features of the energetics and kinetics of the F1-motor. Less

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (27 results)

All 2005 2004 2003 2002 Other

All Journal Article (16 results) Book (3 results) Publications (8 results)

  • [Journal Article] One Rotary Mechanism for F_1-ATPase over ATP Concentrations from Millimolar down to Nanomolar2005

    • Author(s)
      Sakaki N, Shimo-Kon R, Adachi K, Itoh H, Furuike S, Muneyuki E, Yoshida M, Kinosita Jr K.
    • Journal Title

      Biophys J. 88・3

      Pages: 2047-2056

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The Characteristics of the (αV^<371>C)_3(β^<337>C)_<3γ> Double Mutant Subcomplex of the TF_1-ATPase Indicate that the Catalytic Site at the α_<TP>-β_<TP> Interface with Bound MgADP in Crystal Structures of MF_1 Represents a Catalytic Site Containing Inhibitory MgADP2005

    • Author(s)
      Bandyopadhyay, S., Muneyuki, E., Allison, W.S.
    • Journal Title

      Biochemistry 44・7

      Pages: 2441-2448

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] One Rotary Mechanism for F_1-ATPase over ATP Concentrations from Millimolar down to Nanomolar2005

    • Author(s)
      Sakaki N, Shimo-Kon R, Adachi K, Itoh H, Furuike S, Muneyuki E, Yoshida M, Kinosita Jr K.
    • Journal Title

      Biophys J. 88(3)

      Pages: 2047-2056

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The Characteristics of the (αV^<371>C)_3(βR^<337>C)_3γ Double Mutant Subcomplex of the TF_1-ATPase Indicate that the Catalytic Site at the α_<TP>-β_<TP> Interface with Bound MgADP in Crystal Structures of MF_1 Represents a Catalytic Site Containing Inhibitory MgADP2005

    • Author(s)
      Bandyopadhyay, S., Mimeyuki, E., Allison, W.S
    • Journal Title

      Biochemistry 44(7)

      Pages: 2441-2448

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] One Rotary Mechanism for F1-ATPase over ATP Concentrations from Millimolar down to Nanomolar2005

    • Author(s)
      Sakaki N., Shimo-Kon R., Adachi K., Itoh H., Furuike S., Muneyuki E., Yoshida M., Kinosita Jr K.
    • Journal Title

      Biophys J. 88・3

      Pages: 2047-2056

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The Characteristics of the (αV^<371>C)_3(βR^<337>C)_3γ Double Mutant Subcomplex of the TF_1-ATPaae Indicate that the Catalytic Site at the α_<TP>-β_<TP> Interface with Bound MgADP in Crystal Structures of MF_1 Represents a Catalytic Site Containing Inhibitory MgADP2005

    • Author(s)
      Bandyopadhyay, S., Muneyuki, E., Allison, W.S.
    • Journal Title

      Biochemistry 44・7

      Pages: 2441-2448

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Chemo-mechanical coupling in F_1-ATPase revealed by simultaneous observation of nucleotide kinetics and rotation.2004

    • Author(s)
      Nishizaka, T., Oiwa, K., Noji, H., Kimura, S., Muneyuki, E., Yoshida, M., Kinosita, K.Jr.
    • Journal Title

      Nature Structural & Molecular Biology 11

      Pages: 142-148

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Chemo-mechanical coupling in F_1-ATPase revealed by simultaneous observation of nucleotide kinetics and rotation2004

    • Author(s)
      Nishizaka, T., Oiwa, K., Noji, H., Kimura, S., Muneyuki, E., Yashida, M., Kinosita, K.Jr.
    • Journal Title

      Nature Structural & Molecular Biology 11

      Pages: 142-148

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Catalysis and rotation of F_1 motor, cleavage of ATP at the catalytic site occurs in 1ms before 40°substep rotation2003

    • Author(s)
      Shimabukuro, K., Yasuda, R., Muneyuki, E., Hara, K.Y., Kinosita, K.Jr.Yoshida, M.
    • Journal Title

      Proc.Natl.Acad.Sci.USA 100

      Pages: 14731-14736

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Origin of apparent negative cooperativity of F_1-ATPase2003

    • Author(s)
      Sakurako Ono, Kiyotaka Y.Hara, Jun Hirao, Tadashi Matsui, Hiroyuki Noji, Masasuke Yoshida, Eiro Muneyuki
    • Journal Title

      Biochim.Biophys.Acta 1607

      Pages: 35-44

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Evidence for Rotation of V_1-ATPase2003

    • Author(s)
      Hiromi Imamura, Masahiro Nakano, Hiroyuki Noji, Eiro Muneyuki, Shouji Ohkuma, Masasuke Yoshida, Ken Yokoyama
    • Journal Title

      Proc.Natl.Acad.Sci.USA 100

      Pages: 2312-2315

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Catalysis and rotation of F_1 motor ; cleavage of ATP at the catalytic site occurs in 1 ms before 40° substep rotation2003

    • Author(s)
      Shimabukuro, K., Yasuda, R., Muneyuki.E., Hara, K.Y., Kinosita, K.Jr., Yoshida, M.
    • Journal Title

      Proc.Natl.Acad.Sci.USA 100

      Pages: 14731-14736

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cl^- concentration Dependence of the Photovoltage Generation by Halorhodopsin from Halobacterium salinarum2002

    • Author(s)
      Eiro Muneyuki, Chie Shibazaki, Yoichiro Wada, Manabu Yakushizin, Hiroyuki Ohtani
    • Journal Title

      Biophys.J. 83

      Pages: 1749-1759

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] F_1-ATPase changes its conformations upon phosphate release.2002

    • Author(s)
      Masaike T., Muneyuki E., Noji H., Kinosita K.Jr., Yoshida M.
    • Journal Title

      J.Biol.Chem. 277

      Pages: 21643-21649

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] The presence of phosphate at a catalytic site suppresses the formation of the MgADP inhibited form of F_1-ATPase2002

    • Author(s)
      Noriyo Mitome, Sakurako Ono, Toshiharu Suzuki, Katsuya Shimabukuro, Eiro Muneyuki, Masasuke Yoshida
    • Journal Title

      Eur.J.Biochem. 269

      Pages: 53-60

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Cl^-concentration Dependence of the Photovoltage Generation by Halorhodopsin from Halobacterium salinarum2002

    • Author(s)
      Eiro Muneyuki, Chie Shibazaki, Yoichiro Wada, Manabu Yakushizin, Hiroyuki Ohtani
    • Journal Title

      Biophys.J. 83

      Pages: 1749-1759

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] Handbook of ATPase(Chapter 10 Rotation of F_1-ATPase)2004

    • Author(s)
      Muneyuki, E., Yoshida, M.(Eds.M.Futai, Y.Wada, J.Kaplan)
    • Total Pages
      493
    • Publisher
      WILEY-VCH Verlag GmbH & Co.Weinheim
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Book] Handbook of ATPases (Chapter 10 Rotation of F_1-ATPase)2004

    • Author(s)
      Muneyuki, E., Yoshida, M.(Eds.M.Futai, Y.Wada, J.Kaplan)
    • Total Pages
      493
    • Publisher
      WILEY-VCH Verlag GmbH & Co. Weinheim
    • Related Report
      2004 Annual Research Report
  • [Book] シリーズ・ニューバイオフィジックスII-10 生物物理学とはなにか2003

    • Author(s)
      宗行 英朗, 吉田 賢右
    • Total Pages
      308
    • Publisher
      共立出版
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Ono, S., Hara, K.Y., Hirao, J., Matsui, T., Noji, H., Yoshida, M., Muneyuki, E.: "Origin of apparent negative cooperativity of F_1-ATPase"Biochem.Biophys.Acta. 1607. 35-44 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Shimabukuro, K., Yasuda, R., Muneyuki, E., Hara, K.Y., Kinosita, K.Jr., Yoshida, M.: "Catalysis and rotation of F_1 motor ; cleavage of ATP at the catalytic site occurs in 1 ms before 40° substep rotation."Proc.Natl.Acad.Sci.USA. 100. 14731-14736 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Nishizaka T, Oiwa K, Noji H, Kimura S, Muneyuki E, Yoshida M, Kinosita K.Jr.: "Chemomechanical coupling in F_1-ATPase revealed by simultaneous observation of nucleotide kinetics and rotation."Nat.Struct.Mol.Biol.. 11. 142-148 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] 宗行英朗, 吉田賢右: "シリーズニューバイオフィジックスII-10 生物物理学とはなにか 第一章 7 エネルギー変換:ATP合成酵素"曽我部正博, 郷信広編 共立出版. 288 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Mitome, N., Ono, S., Suzuki, T., Shimabukuro, K., Muneyuki, E., Yoshida, M.: "The presence of phosphate at a catalytic site suppresses the formation of the MgADP inhibited form of F_1-ATPase"Eur.J.Biochem.. 269. 53-60 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Masaike T., Muneyuki E., Noji H., Kinosita K.Jr., Yoshida M.: "F_1-ATPase changes its conformations upon phosphate release"J.Biol.Chem.. 277. 21643-21649 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Muneyuki, E., Shibazaki, C., Wada, Y., Yakushizin, M., Ohtani, H.: "Cl^-concentration Dependence of the Photovoltage Generation by Halorhodopsin from Halobacterium salinarum"Biophys.J.. 83. 1749-1759 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Imamura, H., Nakano, M., Noji, H., Muneyuki, E., Ohkuma, S., Yoshida, M., Yokoyama, K.: "Evidence for Rotation of V_1-ATPase"Proc.Natl.Acad.Sci.USA. 100. 2312-2315 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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