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Reactivation of proteins and membrane transport by the heat shock proteins from thermophilic bacteria.

Research Project

Project/Area Number 01870015
Research Category

Grant-in-Aid for Developmental Scientific Research

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionJichi Medical School

Principal Investigator

KAGAWA Yasuo  Jichi Medical School, Facul. Med., Professor, 医学部, 教授 (30048962)

Co-Investigator(Kenkyū-buntansha) NAKANISHI Makoto  Jichi Medical School, Facult. Med., Assistant, 医学部, 助手 (40217774)
OHTA Shigeo  Jichi Medical School, Facult. Med., Assoc. Prof., 医学部, 助教授 (00125832)
HAMAMOTO Toshiro  Jichi Medical School, Facult. Med., Lecturer, 医学部, 講師 (30189625)
平田 肇  自治医科大学, 医学部, 講師 (40049052)
曽根 〓史 (曽根 のぶ史)  自治医科大学, 医学部, 助教授 (20049034)
Project Period (FY) 1989 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥30,100,000 (Direct Cost: ¥30,100,000)
Fiscal Year 1991: ¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1990: ¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1989: ¥17,500,000 (Direct Cost: ¥17,500,000)
Keywordsheat shock protein / thermophilic chaperonin / reconstitution / GroEL / thermophilic gene / hsp gene / ATP synthase / oligomer / synchrotron radiation / chaperonin / biomembrane / protein renaturation / conformationーbuildーup / membrane transport / thermophilic protein / suppression of mutation / 熱ショックタンパク質 / 好熱菌hsp / 高次構造再構成 / シャペロニン / 安定hsp
Research Abstract

Heat shock proteins are essential for tlie formation and restoration of many supramolecular structures. For this purpose, we isolated stable heat shock proteins of 6lkDa, 12kDa and 69kDa, which are similar to GroEL, GroES and DnaK, respectively, 'from the supernatant fraction of thermophilic bacterium PS3 by ATP-Agarose chromatography. Using synthetic DNA of the deduced sequence, the 1.6kbp DNA encoding both proteins was obtained by polymerase chain reaction. The complete sequence of the resulting reading frames showed high. homology to those of the genes encoding GroEL and GroES, and the 12kDa and 6lkDa genes were present in the same operon. 61KDa was also partially similar to the F_1alpha subunit of thermophilic ATP synthase. Using the over-expressed thermophilic F_1alpha subunit it was possible to isolate t
he alpha_3beta_3 and alpha_1beta_1 subunit complexes. These results subunits have been analyzed by synchrotron radiation etc. These results are precious to analyze energy transformation in the reconstituted membrane system.
Patent : Toku-Gan-Hei#2-73664, Heat shock protein, Applicant : Yasuo Kagawa.

Report

(4 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (36 results)

All Other

All Publications (36 results)

  • [Publications] Tamada,H.,Ohta,T.,Hamamoto,T.,Otawara-Hamamoto,Y.,Yanagi,M.,Kagawa,Y.: "Gene structure of heat shock proteins 61KDa and 12KDa (thermphilic chaperonins) of thermophilic bacterium PS3." Biochem.Biophys.Res.Commun. 179. 565-571 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Aloise,P.,Kagawa,Y.and Coleman,P.S.: "Comparative Mg^<2+>-dependent sequential covalent binding stoichiometry of 3'-O-(4-benzoyl)benzoyl adenosine 5'-diphosphate of MF1,TF1,and alpha3-beta3 core complex of TF1." J.Biol.chem.266. 10368-10376 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Harada,M.,Ito,Y.,Sato,M.Aono,O.,Ohta,S.and Kagawa,Y.: "Small-angle X-ray scattering studies of Mg・AT(D)P-induced hexamer to dimer dissociation in the reconstituted alpha3 beta3 complex of ATP synthase from thermophilic bacterium PS3." J.Biol.Chem.266. 11455-11460 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Shirakibara,Y.,Yohda,M.,Kagawa,Y.,Yokoyama,K.and Yoshida,M.: "Purification by dye-ligand chromatography and crystallization study of the F_1-ATPase and its major subunits alpha and beta,from a thermophilic bacterium PS3." J.Biochem.109. 466-477 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 香川 靖雄,玉田 寛,太田 敏子,浜本 敏郎: "好熱菌シャペロニンとATP合成酵素。" Cell Science. 7. 711-717 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 香川 靖雄,太田 成男,浜本 敏郎: "エネルギ-産生膜の形成と機能:シャペロニン類と遺伝子の役割。" 膜. 16. 212-222 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kagawa,Y.: "Biocomputers--The Next Generation from Japan.Bioenergetics and biodevices" Chapman and Hall,New York,Translator:Norman,K., 375 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kagawa,Y.,Ohta,S.,Hamamoto,T.,Harada,M Ito,Y.and Sato,M.: "New Era of Bioenergetics.The alphal betal heterodimer and molecular assembly of ATP synthase" Academic Press,New York,(Mukohata,Y.ed.), 560 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tamada, H., Ohta, T., Hamamoto, T., Otawara - Hamamoto, Y., Yanagi, M., Hiraiwa, H., Hirata, H. and Kagawaw, Y.: "Gene structure of heat shock proteins 61KDa and 12KDa (thermophilic chaperonins) of thermophilic bacterium PS3." Biochemical and Biophysical Research Communications. 179(1). 565-571 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Harada, M., Ito, Y., Sato, M., Aono, O., Ohta, S., and Kagawa, Y.: "Small - angle X - ray scattering studies of Mg・AT (D) P - induced hexamer to dimer dissociation in the reconstituted alpha3 beta3 complex of ATP synthase from thermophilic bacterium PS3." Journal of Biological Chemistry. 266. 11455-11460 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kagawa, Y., Tamada, H., Ohta, T. and Hamamoto, T.: "Thermophilic chaperonin and ATP synthase." Cell Science. 7. 711-717 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kagawa, Y., Ohta, S., Hamamoto, T., Harada, M. Ito, Y., and Sato, M.: Academic Press, N. Y.New Era of Bioenergetics. (Mukohata, Y. ef.)(The alpha1 beta1 heterodimer and molecular assembly of ATP synthase.), (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Tamada,H.,Ohta,T.,Hamamoto,T.,Otawara-Hamamoto,Y.,Yanagi,M.,Kagawa,Y.: "Gene structure of heat shock proteins 61KDa and 12KDa (thermphilic chaperonins) of thermophilic bacterium PS3." Biochem.Biophys.Res.Commun.179. 565-571 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Aloise,P.,Kagawa,Y.and Coleman,P.S.: "Comparative Mg^<2+>-dependent sequential covalent binding stoichiometry of 3'-0-(4-benzoyl) benzoyl adenosine 5'-diphosphate of MF1,TF1,and alpha3-beta3 core complex of TF1." J.Biol.Chem.266. 10368-10376 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Harada,M.,Ito,Y.,Sato,M.Aono,O.,Ohta,S.and Kagawa,Y.: "Small-angle X-ray scattering studies of Mg・AT(D)P-induced hexamer to dimer dissociation in the reconstituted alpha3 beta3 complex of ATP synthase from thermophilic bacterium PS3." J.Biol.Chem.266. 11455-11460 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Shirakibara,Y.,Yohda,M.,Kagawa,Y.,Yokoyama,K.and Yoshida,M.: "Purification by dye-ligand chromatography and crystallization study of the F_1-ATPase and its major subunits alpha and beta,from a thermophilic bacterium PS3." J.Biochem.109. 466-477 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 香川 靖雄、玉田 寛、太田 敏子、浜本 敏郎: "好熱菌シャペロニンとATP合成酵素。" Cell Science. 7. 711-717 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 香川 靖雄、太田 成男、浜本 敏郎: "エネルギ-産生膜の形成と機能:シャペロニン類と遺伝子の役割。" 膜. 16. 212-222 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kagawa,Y.: "Biocomputers--The Next Generation from Japan. Bioenergetics and biodevices" Chapman and Hall,New York,Translator:Norman,K., 375 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kagawa,Y.,Ohta,S.,Hamamoto,T.,Harada,M Ito,Y.and Sato,M.: "New Era of Bioenergetics. The alphal betal heterodimer and molecular assembly of ATP synthase" Academic Press,New York,(Mukohata,Y.ed.), 560 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 香川 靖雄: "Regulation of mitochondrial ATP synthesis in mammalian cells by transcriptional control." Interntl.J.Biochemistry. 22. 219-229 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 香川 靖雄: "Cytosol type electrolyte medium suitable for long term growth of human cells with very low membrane potential." Biochem.Biophys.Res.Commun.169. 1217-1221 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Tomura,H.香川 靖雄: "Novel regulatory enhancer in the gene of the human mitochondrial ATP synthase beta subunit." J.Biol.Chem.265. 6525-6527 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Hayashi,H.: "Effects of ethidium bromide treatment of mouse cells on expression and assembly of nuclearーcoded subunits of complexes involved in oxidative phosphorylation." Biochem.Biophys.Res.Commun.167. 216-221 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ito,Y.: "Smallーangle neutron scattering from the reconstituted TF_1 of H^+ーATPase from thermophilic bacterium PS3 with deuterated subunits." J.Molecular Biol.213. 289-302 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kagawa,Y.: "Lipid metabolism of and energy transduction of cells cultured in serum free synthetic media." Advances in Membrane Fluidity. 8. 91-101 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 香川 靖雄: "岩波講座 分子生物科学7 エネルギ-の生産と運動" 岩波書店、東京, 189 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kagawa,Y.: "In:Bioinstrumentation:Developments and Applications.(Stable oligomeric proteins for biosensors:Protein chemistry,gene engineering and electronics." SpringerーVerlag,New York and Berlin, 1563 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Muneyuki,E.,Kagawa,Y.and Hirata,H.: "Steady state Kinetics of proton translocation catalyzed by thermophilic F_0F_l ATPase reconstituted in planar bilayer membranes." J.Biol.Chem.264. 6092-6096 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Kagawa,Y.,Ohta,s.and Otawara-Hamamoto,Y.: "Alpha 3 beta 3 complex of thermophilic ATP synthase Catalysis without the gamma-subunit." FEBS Letters. 249. 67-69 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yoshimura,H.,Endo,S.,Matsumoto,M.,Nagayama,K.and Kagawa Y.: "Hexagonal structure of two-dimensional crystals of the alpha 3 beta 3 complex of thermophilic ATP synthase." J.Biochem.106. 958-960 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Endo,H.,Hasegawa,K.Narisawa,K.,Tada,K.Kagawa,Y.and Ohta,S.: "Defective gene in lactic acidodis:Abnormal pyruvate Dehydrogenase E_1 a-subunit caused by a frame shift." Am.J.Human Genetics. 44. 358-364 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Tomura,H.,Endo,H.,Kagawa,Y.and Ohta,S.: "Novel regulatory enhancer in the nuclear gene of the human mitochondrial ATP synthase beta-subunit." J.Biol.Chem.265. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Ito,Y.,Harada,M.,Ohta,S.,Kagawa,Y.,Aono,O.,Schefer,J.et al: "Small-angle neutron scattering studies of the reconstituted TF_1 from thermophilic bacterium with deuterated subunits." J.Molecular Biol.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] 香川靖雄: "岩波講座・分子生物科学第7巻エネルギ-の生産と運動" 岩波書店,

    • Related Report
      1989 Annual Research Report
  • [Publications] Kagawa,Y.,Ohta,S.,Hirata,H.,Hamamoto,T.and Karube,I.: "Stable oligomeric proteins for biosensors:Protein chemistry,gene engineering and electronics." Springer-Verlag,Berlin., (1990)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2019-02-15  

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