• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Molecular Mechanisms of Recognition of Target Proteins by the Chaperonin GroEL

Research Project

Project/Area Number 07408017
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionThe University of Tokyo

Principal Investigator

KUWAJIMA Kunihiro  The University of Tokyo ; Graduate School of Science ; Associate Professor, 大学院・理学系研究科, 助教授 (70091444)

Co-Investigator(Kenkyū-buntansha) IKURA Teikichi  The University of Tokyo ; Graduate School of Science ; Research Associate, 大学院・理学系研究科, 助手 (50251393)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥37,000,000 (Direct Cost: ¥37,000,000)
Fiscal Year 1997: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1996: ¥11,400,000 (Direct Cost: ¥11,400,000)
Fiscal Year 1995: ¥21,600,000 (Direct Cost: ¥21,600,000)
KeywordsMolecular chaperone / chaperone / GroEL / Folding mechanism / Globular proteins / alpha-Lactalbumin / Staphylococcal nuclease / シャペロン / スタフィロマッカルヌクレアーゼ
Research Abstract

For the purpose of understanding the relationship between the protein refolding in vitro and the protein folding in a biological cell, we studied the effect of the chaperonin GroEL on the refolding kinetics of alpha-lactalbumin (alphaLA) and staphylococcal nuclease (SNase) by stopped-flow fluorescence spectroscopy. The results have shown that the effect of GroEL on the refolding reaction is apparently very different for SNase and apo-alphaLA.When the apparent refolding rate was estimated by measurements of the refolding reaction at different concentrations of GroEL,the refolding rate constant was changed in alphaLA,while the amplitude of the major kinetic process of the free refolding was decreased in SNase without large changes in the rate constants of the individual processes, and only the slow refolding process that occurred in the GroEL-bound state was observed in excess GroEL.From ionic-strength dependence of the refolding reaction in the presence of GroEL,the above difference bet … More ween the two target proteins was shown to be due to a difference in the electrostatic properties of the proteins. alphaLA is a acidic protein having a net charge of -7 at neutral pH while SNase is a basic protein with a net charge of +12. On the other hand, GroEL is a strongly acidic protein having a charge of -20 per monomer (-280 per 14mer). Therefore, there must be electrostatic repulsion between alphaLA and GroEL and attraction between SNase and GroEL.From the present study, it is concluded that the long-range electrostatic interactions as well as the hydrophobic interactions are important for the recognition of a target protein by GroEL.
Next, we simulated the refolding processes of a protein under the influence of GroEL on a computer, on the basis of a scheme that the target protein is reversibly bound to GroEL but can also refold in the GroEL-bound state. The results have shown that although the effects of GroEL on the refolding reactions of the above two target proteins are apparently very different, they both can be interpreted in terms of the same unified reaction scheme. Less

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (50 results)

All Other

All Publications (50 results)

  • [Publications] Katsumata, Kumiko: "Dominant forces in the recognition of a transient folding intemediate ofα-lactalbumin by GroEL" J.Mol.Biol.264. 643-649 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, Kunihiro: "Stopped-flow circular dichroism" In Circular dichroism and the conformational analysis of biomolecules(Fasman,G.D.ed). 159-182 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kataoka, Mikio: "Structural characterization of the molten globule of α-lactalbumin by solution X-ray scattering" Protein Science. 6. 422-430 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Okazaki, Akira: "Hydrogen-exchage kinetics of reduced α-lactalbumin bound to the chaperonin GroEL" J.Biochem.(Tokyo). 121. 534-541 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ikura, Teikichi: "Kinetic folding and cis/trans prolyl isomerization of staphylococcal nuclease.A study by stopped-flow absorption.stopped-flow circular dichroism and molecular dynamics simulations" Biochemistry. 36. 6529-6538 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Arai, Munehito: "Kinetic Refolding of β-Lactoglobulin.Studies by Synchrotron X-Ray Scattering,and Circular Dichroism,Absorption and Eluorescence Spectroscopy" J.Mol.Biol.275. 149-162 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsurupa, Galina P.: "Refolding Kinetics of Staphylococcal Nuclease and Its Mutants in the Presence of the Chaperonin GroEL" J.Mol.Biol.(In the press.). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, Kunihiro: "Molten globule" In The Encyclopedia of MolecularBiology(Creighton,T.E.ed). (In the press.). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kalnin, Nikolay N.: "Kinetic folding and unfolding ofstaphylococcal nuclease and its six mutants studied by stopped-flow circular dichroism" Proteins:Structure,Function & Genetics. 23. 163-176 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Uchiyama, Hidefumi: "Effects of amino acid substitutions in the hydrophobic core of α-lactalbumin on the stability of the molten globule state" Protein Eng.8. 1153-1161 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, Kunihiro: "Circular dichroism" In Protein stability and folding. 115-135 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, Kunihiro: "The molten globule state of α-lactalbumin" FASEB J.10. 102-109 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Arai, Munehito: "Rapid formation of a molten globule intermediate in refolding of α-lactalbumin" Folding & Design. 1. 275-287 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Katsumata, Kumiko: "Effect of GroEL on the refoldingkinetics of α-lactalbumin" J.Mol.Biol.258. 827-838 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Semisotnov, Gennady V.: "Protein globularization during folding.A study by synchrotron small-angle X-ray scattering" J.Mol.Biol.262. 559-574 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, Kunihiro: "The burst-phase intermediate in the refolding of β-lactoglobulin studied by stopped-flow circular dichroism and absorption spectroscopy" J.Mol.Biol.264. 806-822 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ikaguchi, Masamichi: "Transition State in the Folding of α-Lactalbumin Probed by the 6-120Disulfide Bond" Protein Science. (In the press.). (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kalnin, N.N.and Kuwajima, K.: "Kinetic folding and unfolding of staphylococcal nuclease and its six mutants studied by stopped-flow circular dichroism" Proteins. 23. 163-176 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Uchiyama, H. ; Perez-Prat, E.M. ; Watanabe, K.Kumagai, I.and Kuwajima, K.: "Effects of amino acid substitutions in the hydrophobic core of alpha-lactalbumin on the stability of the molten globule state" Protein Eng.8. 1153-1161 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, K.: "Circular dichroism" In Protein stability and folding (Methods Mol.Biol.Vol.40) (Shirley, B.A.ed) 115-135, Humana Press Inc.Totowa, NJ. (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, K.: "The molten globule state of alpha-lactalbumin." FASEB J.10. 102-109 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Arai, M and Kuwajima, K.: "Rapid formation of a molten globule intermediate in refolding of alpha-lactalbumin." Folding & Design. 1. 275-287 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Katsumata, K. ; Okazaki, A.and KuwaJima, K.: "Effect of GroEL on the refolding kinetics of alpha-lactalbumin" J.Mol.Biol.258. 827-838 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Semisotnov, G.V. ; Kihara, H. ; Kotova, N.V. ; Kimura, K. ; Amemiya Y. ; Wakabayashi, K. ; Serdyuk, I.N. ; Timchenko, A.A. ; Chiba, K. ; Nikaido, K. ; Ikura, T.and Kuwajima, K.: "Protein globularization during folding. A study by synchrotron small-angle X-ray scattering." J.Mol.Biol.262. 559-574 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, K. ; Yamaya, H.and Sugai, S.: "The burst-phase intermediate in the refolding of beta-lactoglobulin studied by stopped-flow circular dichroism and absorption spectroscopy" J.Mol.Biol.264. 806-822 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Katsumata, K. ; Okazaki, A. ; Tsurupa, G.P.and Kuwajima, K.: "Dominant forces in the recognition of a transient folding intermediate of alpha-lactalbumin by GroEL" J.Mol.Biol.264. 643-649 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, K.: "Stopped-flow circular dichroism" In Circular dichroism and the conformational analysis of biomolecules (Fasman, G.D.ed) pp.159-182, Plenum, New York.(1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kataoka, M. ; Kuwajima, K. ; Tokunaga, F.and Goto, Y.: "Structural characterization of the molten globule of alpha-lactalbumin by solution X-ray scattering." Protein Sci.6. 422-430 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Okazaki, A. ; Katsumata, K.and Kuwajima, K.: "Hydrogen-exchange kinetics of reduced alpha-lactalbumin bound to the chaperonin GroEL" J.Biochem. (Tokyo). 121. 534-541 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ikura, T. ; Tsurupa, G.P.and Kuwajima, K.: "Kinetic folding and cis/trans prolyl isomerization of staphylococcal nuclease. A study by stopped-flow absorption, stopped-flow circular dichroism and molecular dynamics simulations." Biochemistry. 36. 6529-6538 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Arai, M. ; Ikura, T. ; Semisotnov, G.V. ; Kihara, H. ; Amemiya, Y.and Kuwajima, K.: "Kinetic Refolding of beta-Lactoglobulin. Studies by Synchrotron X-Ray Scattering, and Circular Dichroism, Absorption and Fluorescence Spectroscopy." J.Mol.Biol.275. 149-162 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsurupa, G.P. ; Ikura, T. ; Makio, T.and Kuwajima, K.: "Refolding Kinetics of Staphylococcal Nuclease and Its Mutants in the Presence of the Chaperonin GroEL" J.Mol.Biol.(In the press.). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kuwajima, K.: "Molten globule." In The Encyclopedia of Molecular Biology (Creighton T.E.ed) Wiley. (In the press.). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Ikeguchi, M. ; Fujino, M. ; Kato, M. ; Kuwajima, K.And Sugai, S.: "Transition State in the Folding of alpha-Lactalbumin Probed by the 6-120 Disulfide Bond." Protein Sci.(In the press.). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Tsurupa,GP: "Refolding Kinetics of Staphylococcal Nuclease and Its Mutants in the Presence of the Chaperonin GroEL" J.Mol.Biol.(In the press).

    • Related Report
      1997 Annual Research Report
  • [Publications] Arai,M: "Kinetic Refolding of β-Lactoglobulin.Studies by Synchrotron X-Ray Scattering,and Circular Dichroism,Absorption and Fluoressence Spectroscopy" J.Mol.Biol.275. 149-162 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ikura,T: "Kinetic folding and cis/trans prolyl isomerization of staphylococcal nuclease.A study by stopped-flow absorption,stopped-flow circular dichroism and molecular dynamics simulations" Biochemistry. 36. 6529-6538 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Okazaki,A: "Hydrogen-exchange kinetics of reduced α-lactalbumin bound to the chaperonin GroEL" J.Biochem.(Tokyo). 121. 534-541 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kataoka,M: "Structual characterization of the molten globule of α-lactalbumin by solution X-ray scattering" Protein Sci.6. 422-430 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Okazaki,A: "Hydrogen-Exchange Kinetics of Reduced-alpha-Lactalbumin Bound to the Chaperonin GroEL" J.Biochem.(Tokyo). in press (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kuwajima,Kunihiro: "The Burst-Phase Intermediate in the Refolding of beta-Lactoglobulin Studied by Stopped-Flow Circular Dichroism and Absorption Spectroscopy" J.Mol.Biol.264. 806-822 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Katsumata,K.: "Dominant Forces in the Recognition of a Transient Folding Intermediate of alpha-Lactalbumin by GroEL" J.Mol.Biol.264. 643-649 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Semisotnov,G.V.: "Protein Globularization During Folding.A study by Synchrotron Small-Angle X-Ray Scattering" J.Mol.Biol.262. 559-574 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Arai,M.: "Repid formation of a molten globule intermediate in refolding of alpha-lactalbumin" Folding & Design. 1. 275-287 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Katsumata,K.: "Effect of GroEL on the refolding kinetics of alpha-lactalbumin" J.Mol.Biol.258. 827-838 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Katsumata,Kumiko: "Effect of GroEL on the refolding kinetics of alpha-lactalbumin" J.Mol.Biol.(in Press). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kuwajima,Kunihiro: "The molten globule state of alpha-lactalbumin" FASEB J.10. 102-109 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Uchiyama,Hidefumin: "Effect of amino acid substitution in the hydrophobic core of alpha-lactalbumin on the stability of the molten globule state" Protein Eng.(in Press). (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kalnin,Nikolay N.: "Kinetic folding and unfolding of staphylococcal nuclease and its six mutants studied by stopped-flow circular dichroism" Proteins:Structure,Functions and Genetics. 23. 163-176 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 桑島邦博: "GroELによる標的タンパク質認識の分子機構" 蛋白質核酸酵素. (印刷中). (1996)

    • Related Report
      1995 Annual Research Report

URL: 

Published: 1995-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi