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Effective temperature of proteins in the active states

Research Project

Project/Area Number 12304022
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field 物理学一般
Research InstitutionOsaka University

Principal Investigator

YANAGIDA Toshio  Graduate School of Frontier Biosciences, Professor, 生命機能研究科, 教授 (30089883)

Co-Investigator(Kenkyū-buntansha) IWANE Atsuko  Graduate School of Frontier Biosciences, Assistant Professor, 生命機能研究科, 助手 (30252638)
SEKIMOTO Ken  Institute of Basic Physics, Kyoto University, Professor, 基礎物理学研究所, 教授 (00179342)
Project Period (FY) 2000 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥47,300,000 (Direct Cost: ¥41,300,000、Indirect Cost: ¥6,000,000)
Fiscal Year 2002: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
Fiscal Year 2001: ¥15,600,000 (Direct Cost: ¥12,000,000、Indirect Cost: ¥3,600,000)
Fiscal Year 2000: ¥21,300,000 (Direct Cost: ¥21,300,000)
Keywordschemi-mechano coupling / thermal ratchet / Browninan movement / effective temperature / rotational elasticity / molecular motors / actomyosin / 科学-力学共役 / アクチン / ミオシン / 分子モータ / 蛋白質 / 光ピンセット
Research Abstract

Many proteins work using chemical energy that generates during hydrolysis of ATP into ADP and P1. A basic question, how proteins converts this chemical energy to mechanical works remains unsolved. The aims of this study were to examine if the effective temperature of a protein molecule excited by chemical energy of ATP is really increased and. If so, to construct a model to explain how chemical and mechanical energies are converted by a protein molecule. We chose actomyosin as the experimental system, and during the period of this grant, we obtained the following achievements.
(1) We have developed an experimental system to measure the effective temperature of an actin filament interacting with myosin molecules: an actin filament was horizontally held in solution being caught two microbeads attached to the both ends using double beam optical tweezers. Relative rotational Brownian movement of the two beads was measured and the effective temperature was calculated from the mean square of the angle between the two beads and the relaxation tirne.
(2) We have measured the rotational fluctuation of an actin filament interacting with myosin in the presence of ATP: Myosin VI, which has a 36nm step length per one ATP hydrolysis, rotated 45 degree during five 36nm steps. This movement cannot be explained according to simple walking model along the coiled actin filament.
(3) Myosin molecules are diffusing along the thermal ratchet of an actin filament. The ratio of forward and backward steps was measured using singlemolecule technique. The result indicates that difference of the energy barrier for forward and backward direction is 3kBT. This value is almost same for another protein motor kinesin.

Report

(4 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] S.Nishikawa, K.Homma, Y.Komori et al.: "Class VI myosin moves processively along actin filament backwards with large steps"Biochem. Biophys. Res. Commun.. 290. 311-317 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H.Tanaka, K.Homma, A.H.Iwane et al.: "The motor doamin determines the large step of myosin-V"Nature. 415. 192-195 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M.Nishiyama, H.Higuchi, T.Yanagida: "Chemomechanical coupling of the ATPase cycle to the forward and backward movements of single kinesin molecules"Nature Cell Biology. 4. 790-797 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y.Inoue, A.H.Iwane, T.Miyai, E.Muto, T.Yanagida: "Motility of single one-headed kinesin molecules along microtubles"Biophysical Journal. 81. 2838-2850 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M.Nishiyama, E.Muto, Y.Inoue, T.Yanagida, H.Higuchi: "Substeps within the 8-nm step of the ATPase cycle of single kinesin molecules"Nature Cell Biology. 3. 425-428 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Yanagida, A.H.Iwane: "A large step for myosin"Proc. Natl. Acad. Sci. USA. 97. 9357-9359 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Yanagida, H.Tanaka, K.Kitamura, T.Wazawa, et al.: "Single molecule techniques in biophysics. "Na/K-ATPase and Related ATPases""ELSEVIER. (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] S, Nishikawa, K, Homma, Y, Komori: "Class VI myosin moves pcocessively along actin filament backwards with large steps"Biochem. Biophys. Res. Commun. 290. 311-317 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H, Tanaka, K, Homma, A, H, Iwane: "The motor doamin determines the large step of myosin-V"Nature. 415. 192-195 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M, Nishiyama, H, Higuchi, T, Yanagida: "Chemomechanical coupling of the ATPase cycle to the forward and backward movements of single kinesin molicules"Nature Cell Biol.. 4. 790-797 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y, Inoue, A, H, Iwane, T, Miyai, E, Muto, T, Yanagida: "Motility of single one-headed kinesin molecules along microtubles"Biophys. J. 81. 2838-2850 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] M, Nishiyama, E, Muto, Y, Inoue, T, Yanagida, H, Higuchi: "Substeps within the 8-nm step of the ATPase cycle of single kinesin mdlecules"Nature Cell Biol.. 3. 425-428 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T, Yanagida, A, H, Iwane: "A large step for myosin"Proc. Natl. Acad. Sci. USA.. 97. 9357-9359 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T, Yanagida, H, Tanaka, K, Kitamura, T, Wazawa: "Single molecule techniques in biophysics"Na/K-ATPase and related ATPases. ELSEVIER. 71-85 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.ide, Y.Takeuchi, T.Yanagida: "Development of an Experimental Apparatus for Simultaneous Observation of Optical and Electrical Signals from Single Ion Channels"Single Molecules. 3. 33-42 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] S.Nishikawa, K.Homma, Y.Komori et al.: "Class VI myosin moves processively along actin filament backwards with large steps"Biochem. Biophys. Res. Commun.. 290. 311-317 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] H.Tanaka, K.Homma, A.H.Iwane et al.: "The motor doamin determines the large step of myosin-V"Nature. 415. 192-195 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] M.Nishiyama, H.Higuchi, T.Yanagida: "Chemomechanical coupling of the ATPase cycle to the forward and backward movements of single kinesin molecules"Nature Cell Biology. 4. 790-797 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ide, T., Takeuchi, Y., Aoki, T., Yanagida, T.: "Simultaneous optical and electrical recording of a single ion-channel"Jpn J Physiol.. 52. 429-434 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nishiyama, M.et al.: "Substeps within the 8-um step of the ATPase cycle of single kinesin molecules"Nature Cell Biology. 3. 425-428 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Y.Ishii et al.: "Single molecule nanomanipulation of bio molcules"Trends in Biotechnology. 19. 211-216 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Ishijima, A., Yanagida T.: "Single molecule nano-biosaience"Trends in Biochemical Siences. 26. 438-444 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Inoue, Y.et al.: "Motility of single one-headed kinesin molecules along microtables"Biophysical Journal. 81. 2838-2850 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Aoki, T.et al.: "Non-contact electrostatic surface force imaging of single protein filaments using intermolecular force microscopy"Single Molecules. 2. 183-190 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Ueda, M.et al.: "Single molecule analysis of chemotactic signaling in Dictyostellum cells"Science. 294. 864-867 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] T.Yanagida,K.Kitamura,H.Tanaka,A.H.Iwane & S.Esaki: "Single molecule analysis of the actomyosin motor"Current Opinion in Cell Biology. 12. 20-25 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Wazawa,Y.Funatsu & T.Yanagida: "Spectral fluctuation of a single fluorophore conjugated to a protein molecule"Biophys.J.. 78. 1561-1569 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Y.Sako,S.Minoguchi & T.Yanagida: "Single-molecule imaging of EGFR signalling on the surface of living cells"Nature Cell Biology. 2. 168-172 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Y.Ishii,Y.Kimura,K.Kitamura,H.Tanaka,T.Wazawa & T.Yanagida: "Imaging and Nano-manipulation of Single Actomyosin Motors at Work"Clinical and Experimental Pharmacology and Physiology. 27. 229-237 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] T.Yanagida & A.H.Iwane: "A large step for myosin"PNAS.. 97. 9357-9359 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Y.Sako,K.Hibino,T.Miyauchi,Y.Miyamoto,M.Ueda & T.Yanagida: "Single-Molecule Imaging of Signaling Molecules in Living Cells"Single Molecules. 1. 159-163 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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