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Structure and function of NaィイD1+ィエD1-pumping NADH-quinone reductase

Research Project

Project/Area Number 09672213
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionChiba University

Principal Investigator

UNEMOTO tsutomu  Chiba University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (30089601)

Co-Investigator(Kenkyū-buntansha) HAYASHI Maki  Chiba University, Faculty of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (50092086)
Project Period (FY) 1997 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1999: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsSodium ion pump / NADH-quinone reductase / nqr gene / Respiratory chain / Vibrio alginolyticus / Marine bacterium / nqp gene / Marine bacterium / Na^+ポンプ / NADH-キノンリダクターゼ / 呼吸鎖 / ナトリウム輸送 / 海洋細菌 / コロルミシン / Na^+-translocating / vespiratery chain / Na^+ pump / Vivric alginalyticus / marine bacteria / over expression
Research Abstract

Earlier studies in this laboratory demonstrated that the NaィイD1+ィエD1-translocating NADH-quinone reductase (NQR) of the marine bacterium Vibrio alginolyticus is composed of 6 structural genes. In this studies, 6 subunits corresponding to Nqr1, Nqr2, Nqr3, Nqr4, Nqr5 and Nqr6 could be detected on SDS-PAGE by modifying the detection method of proteins. The open reading frame of each subunit was identified from its N-terminal amino acid sequence. The genetic defect of a mutant Nap1 was found to be caused by the insertion of a 1.2kbp DNA fragment into the C-terminal region of nqr6 gene. We dissociated the purified NQR complex to subunit components, Nqr1, Nqr3, Nqr6 and Nqr2,4,5. The latter Nqr2,4,5 fraction contained all hydrophobic subunits and this fraction was difficult to further dissociate into each component subunit. We could construct a reconstitution system starting from these subunits, and it was found that all six subunits are essential for the NQR activity. Recently, a novel antibiotic, korormicin, was found to specifically inhibit NQR activity. The mode of korormicin inhibition was purely nonncompetitive for Q-1 with the inhibitor constant of 82 pM. The site of korormicin binding was partially overlapped with that of HQNO.

Report

(4 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • 1997 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] M.Hayashi et al.: "Genetic defect of the sodium pump-defective mutant Nap1 from the marine"Biochem. Molecular Biol. Intern.. 41. 41-47 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y.Nakayama et al.: "Identification of six subunits constituting Na^+_-translocating NADH-quinone"FEBS Lett.. 422. 240-242 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K.Yoshikawa et al.: "Korormicin, an antibiotic specific for Gram-negative marine bacteria,"J. Antibiotic.. 52. 182-185 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Y.Nakayama et al.: "Inhibitor studies of a new antibiotic, korormicin, HQNO and Ag^+ toward"Biol Pharm. Bull.. 22. 1064-1067 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] 畝本力: "海洋細菌の生体エネルギー論―呼吸鎖に共役したNa^+ポンプ"YAKUGAKU ZASSHI. 120. 16-27 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Hayashi,M., Fujii,J. and Unemoto,T.: "Genetic defect of the sodium pump-defective mutant Nap1 from the marine Vibrio alginolyticus"Biochem. Molecular Biol. Intern. 41. 41-47 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakayama,Y., Hayashi,M. and Unemoto T.: "Identification of six subunits constituting NaィイD1+ィエD1-translocating NADH-quinone reductase from the marine Vibrio alginolyticus"FEBS Lett.. 422. 240-242 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yoshikawa,K., Nakayama,Y., Hayashi,M.., Unemoto,T. and Mochida,K.: "Korormicin, an antibiotic specific for Gram-negative marine bacteria, strongly inhibits the respiratory chain-linked NaィイD1+ィエD1-translocating NADH-quinone reductase from the marine Vibrio alginolyticus"J.Antibiotic. 52. 182-185 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Nakayama,Y., Hayashi,M., Yoshikawa,K., Mochida,K. and Unemoto,T.: "Inhibitor studies of a new antibiotic, korormicin, HQNO and AgィイD1+ィエD1toward the NaィイD1+ィエD1-translocating NADH-quinone reductase from the marine Vibrio alginolyticus"Biol. Pharm. Bull. 22. 1064-1067 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Unemoto,T.: "Bioenergetics of marine bacteria-respiratory chain-coupled NaィイD1+ィエD1 pump (in Japanese)"YAKUGAKU ZASSHI. 120. 16-27 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] K. Yoshikawa et al.: "Korormicin, an Antibiotic Specific for Gram-Negative Marine Bacteria, ・・・・・・・・・・・"J. Antibiotic.. 52. 182-185 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Y. Nakayama et al.: "Inhibitor Studies of a New Antibiotic, Korormicin, HQNO and Ag^+ toward ・・・・・・・・・・"Biol. Pharm. Bull.,. 22. 1064-1067 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 畝本 力: "海洋細菌の生体エネルギー論-呼吸鎖に共役したNa^+ポンプ-"YAKUGAKU ZASSHI. 120. 16-27 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] K.Yoshikawa et al.: "Korormicin,an Antibiotic Specific for Gram-negative Marine Bacteria,Strongly inhibit Respiratory Chain-linked Na^+-Translocating NADH:Quinone Reductase from the Marine vibrio alginolyticus." The Journal of Antibiotics. 52・2. 182-185 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] M.Hayashi, J.Fujii and T.Unemoto: "Genetic defect of the sodium pump-defective mutant Nap-1 from the manne Vibrio alginolyticus." Biochemistry and Molecular Biology International. 41・1. 41-47 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Nakayama, M.Hayashi and T.Unemoto: "Identification of six subunits constituting Na_+-translocating NADH-quinone reductase from the marine Vibrio algynolyticus." FEBS Letters. 422. 240-242 (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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