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Molecular Mechanism and Structural Basis of Ubiquinone-Redox Reaction in Bacterial Respiratory Chains

Research Project

Project/Area Number 12460045
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionYamaguchi University

Principal Investigator

MATSUSHITA Kazunobu  Faculty of Agriculture, Yamaguchi University, Professor, 農学部, 教授 (50107736)

Co-Investigator(Kenkyū-buntansha) YAMADA Mamoru  Faculty of Agriculture, Yamaguchi University, Professor, 農学部, 教授 (30174741)
MOGI Tateshi  Graduate school of Science, Tokyo University, Associate Professor, 大学院・理学研究科, 助教授 (90219965)
MIYOSHI Hideto  Graduate school of Agriculture, Kyoto University, Associate Professor, 大学院・農学研究科, 助教授 (20190829)
TOYAMA Hirohide  Faculty of Agriculture, Yamaguchi University, Associate Professor, 農学部, 助教授 (60240884)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 2001: ¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 2000: ¥8,800,000 (Direct Cost: ¥8,800,000)
KeywordsAzidoquinone / Photo-affinity labeling / Ubiquinone / Escherichia coli Glucose dehydrogenase / Alcohol Dehydrogenase of acetic acid bacteria / Bacterial respiratory chain / 光親和性標識法 / 大腸菌チトクロムbo / 呼吸鎖
Research Abstract

In this research project, search for ubiquinone-binding site and molecular mechanism of redox reaction of ubiquinone was carried out in several ubiquinone-reacting enzymes in the respiratory chains of Escherichia coli and Gluconobacter suboxydans. The objective ubiquinone-reacting enzymes were cytochrome bo oxidase (Cyo), glucose dehydrogenase (GDH), and succinate dehydrogenases (SDH) of E. coli, and alcohol dehydrogenases (ADH) of G. suboxydans. In order to do this research, detection of the bound quinone in the enzymes and also photo-affinity labeling with azidoquinone of the enzymes followed by analysis of the labeled subunit and further peptides by MALDI-TOF-MS were mainly performed.
1) 2-azido-Q2 and also 3-azido-Q2 were successfully synthesized via the precursors, 2- and 3- metylated quinone compounds, and also the azido-moiety was found to be decomposed by UV irradiation. 2) Labeling conditions for Cyo of E. coli with both 2-azido- and 3-azido-Q2 were established. 3) Ubiquinone-reacting site of E. coli GDH was shown to be present in the C- terminal hydrophilic PQQ domain, which could be done by preparing the C-terminal domain by site-directed mutagenesis and the reactivity toward ubiquinone. 4) In E. coil SDH, ubiquinone-reacting site was shown to be present in the membrane-anchoring subunit by photo-affinity labeling with azidoquinone. 5) When purified G. suboxydans ADH with dodecyl maltoside, the enzyme retained tightly ubiquinone-10 inside the molecule, and also shown to be as the ubisemiquinone form. 6) In G. suboxydans ADH, the subunit II was shown to be labeled with azidoquinone, especially the ubiquinone reduction site, but not the ubiquinol oxidation site. And also the labeled quinone was found to work as electron mediator to pooled ubiquinone.

Report

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

    (51 results)

All Other

All Publications (51 results)

  • [Publications] Uchida, T.: "Resonance Raman studies of oxo intermediates in the reaction of pulsed cytochrome bo with hvdrogen peroxide"Biochemistry. 39. 6669-6678 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi, K.: "Transient tormation of ubisemiquinone radical and subsequent electron transfer processes in the Eseherichia colicytochrome bo revealed by pulse radiolysis"Biochemistry. 39. 15620-15625 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tsubaki, M.: "Probing molecular structure of dioxygen reduction site of bacterial quinol oxidases through ligand binding to the redox metal centers"J.Inorg.Biochem.. 82. 19-25 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Elias, MD: "Functions of amino acid residues in the active site of Escherichia coli PQQ-containing quinoprotein glucose dehydrogenase"J.Biol.Chem.. 275. 7321-7326 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Miyadera, H.: "Altered quinone biosynthesis in the long-kived clk-lmutants of Caenorhabditis elegans"J.Biol.Chem.. 276. 7713-7716 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ohmura, S.: "Anthelmintic compound, nafuredin, shows selective inhibition of complex I in helminth mitochondria"Proc.Natl.Acad.Sci.USA. 98. 60-62 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] O.Adachi: "Membrane-bound quinoprotein D-arabitol dehydrogenase of Gluconobacter suboxydans IFO 3257:A versatile enzyme for the oxidative fermentation of various ketoses"Biosci.Biotechnol.Biochem.. 65. 2755-2762 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Miyoshi, H: "Probing the ubiquinone reduction site in bovine mitochondrial complex I using a series of synthetic ubiquinones and inhibitors"J.Bioenerg.Biomembr. 33. 223-231 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mitani, S: "The biochemica mode of action of the novel selective fungkide cyazofamid : specific inhibition of mitochondrial complex III in Pythium spinosum"Pestic.Biochem.Physiol.. 71. 107-115 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K.Matsushita: "NADH dehydrogenase of Corynebacterium glutamicum. Purification of an NADH dehydrogenase II homolog able to oxidize NADPH"FEMS Microbiol.Lett.. 204. 271-276 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] MD Elias: "C-terminal periplasmic domain of Escherichia coil Quinoprotein glucose dehydrogenase transfers electrons to ubiquinone"J.Biol.Chem.. 276. 48356-48361 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K.Matsushita: "Quinoproteins : structure, function and biotechnological applications"Appl.Micriobiol.Biotechnol.. 58. 13-22 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K.Sakamoto: "Synthesis and Electron-transfer Activity of Azido Ubiquinone-2"J.Pesticide Sci.. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] H.Miyadera: "Quinones in long-lived clk-1 mutants of Caenorhabditis elegans"FEBS Lett.. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 茂木立志: "解き明かされた大腸菌の呼吸の仕組み"蛋白質核酸酵素. 47(印刷中). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Matsushita, K.: "Biochemistry and Molecular Biology of vitamin B6 and PQQ-dependent proteins"Birkhauser Verlag AG. 377 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 茂木立志: "細胞呼吸の膜エネルギー装置:起源と分子進化"「生体膜のエネルギー装置(吉田賢右・茂木立志編)」共立出版. 21 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 松下一信: "廣川タンパク質化学 第4巻酵素 4.1オキシドレダクターゼ"廣川書店. 353 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Uchida T, T. Mogi, and T. Kitagawa: "Resonance Raman studies of oxo intermediates in the reaction of pulsed cytochrome bo with hydrogen peroxide"Biochemistry. 39. 6669-6678 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kobayashi K, S. Tagawa, and T, Mogi: "Transient formation of ubisemiquinone radical and subsequent electron transfer process in the Escherichia coli cytochrome bo revealed by pulse radiolysis"Biochemistry. 39. 15620-15625 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Tsubaki M, H. Hori, and T. Mogi: "Probing molecular structure of dioxygen reduction site of bacterial quinol oxidases through ligand binding to the redox metal centers"J. Inorg. Biochem. 82. 19-25 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Elias MD, M. Tanaka, H. Izu, K. Matsushita, O. Adachi, & M. Yamada: "Functions of amino acid residues in the active site of Escherichia coli PQQ-containing quinoprotein glucose dehydrogenase"J. Biol. Chem. 275. 7321-7326 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Miyadera H, Amino H, Hiraishi A, Taka H, Murayama K, Miyoshi H, Sakamoto K, Ishii N, Hekimi S, and Kita K: "Altered quinone biosynthesis in the long-lived clk-I mutants of Caenorhabditis. elegans"J. Biol. Chem. 276. 7713-7716 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ohmura S, Miyadera H, Ui H, Shiomi K, Yamaguchi Y, Masuma R, Nakamitsu T, Takano D, Sunazuka T, Harder A, Kolbl H, Namikoshi M, Miyoshi H, Sakamoto K, and Kita K: "Anthelmintic compound, nafuredin, shows selective inhibition of complex I in helminth mitochondria"Proc. Natl. Acad. Sci. USA. 98. 60-62 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Miyoshi H: "Probing the ubiquinone reduction site in bovine mitochondrial complex I using a series of synthetic ubiquinones and inhibitors"J. Bioenerg. Biomembr. 33. 223-231 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Mitani S, Araki S, Takii Y, Ohshima T, Matsuo N, and Miyoshi H: "The biochemical mode of action of the novel selective fungicide cyazofamid : specific inhibition of mitochondrial complex III in Pythium spinosum"Pestic. Biochem. Physiol. 71. 107-115 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K. Matsushita, A.Otofuji, M. Iwahashi, H. Toyama & O. Adachi: "NADH dehydrogenase of Coryneacterium glutamicum. Purification of an NADH dehydrogenase II homolog able to oxidize NADPH"FEMS Microbiol. Lett. 204. 271-276 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Elias MD, Tanaka M, Sakai M, Toyama H, Matsushita K, Adachi O, Yamada M: "C-terminal periplasmic domain of Escherichia coli Quinoprotein glucose dehydrogenase transfers electrons to ubiquinone"J. Biol. Chem. 276. 48356-48361 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] O. Adachi, Y, Fujii, M. F. Ghaly, H. Toyama E. Shinagawa, & K. Matsushita: "Membrane-bound quinoprotein D-arabitol dehydrogenase of Gluconobacter suboxydans IFO 3257 : A versatile enzyme for the oxidative fermentation of various ketoses"Biosci. Biotechnol. Biochem. 65. 2755-2762 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K. Matsushita, H. Toyama, M. Yamada, and O. Adachi: "Quinoproteins : structure, function, and biotechnological applications"Appl. Micriobiol. Biotechnol. 58. 13-22 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Miyadera H, Kano K, Miyoshi H, Ishii N, Hekimi S, and Kita K: "Quinones in long-lived clk-1 mutants of Caenorhabaditis elegans"FEBS Lett. 512. 33-37 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K. Sakamoto, K. Nomura, H. Miyoshi: "Synthesis and Electron-transfer Activity of Azido ubiquinone-2"J. Pesticide Sci. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] T. Mogi: "Solved mechanism of Escherichia coli respiration"Protein, Nucleic Acid, and Enzyme. 47 (in press) (in Japanese). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Matsushita K, T. Yamashita, N. Aoki, H. Toyama, O. Adachi: "Electron transport systems for quinohemoprotein type II alcohol dehydrogenase of Pseudomonas putida HK5. Biochemistry and Molecular Biology of vitamin B6 and PQQ-deoendent proteins"Birkhauser Verlag AG. 219-224 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ohmura, S.: "Anthelmintic compound, nafuredin, shows selective inhibition of complex I in helminth mitochondria"Proc.Natol.Acad.Sci.USA. 98. 60-62 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Miyoshi, H.: "Probing the ubiquinone reduction site in bovine mitochondrial complex I using a series of synthetic ubiquinones and inhibitors"J.Bioenerg. Biomembr. 33. 223-231 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Mitani, S: "The biochemical mode of action of the novel selective fungicide cyazofamid : specific inhibition of mitochondrial complex III in Pythium spinosum"Pestic. Biochem. Physiol.. 71. 107-115 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Matsushita: "NADH dehydrogenase of Corynebacterium glutamicum. Purification of an NADH dehydrogenase II homolog able to oxidize NADPH"FEMS Microbiol. Lett.. 204. 271-276 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] MD Elias: "C-terminal periplasmic domain of Escherichia coli Quinoprotein glucose dehydrogenase transfers electrons to ubiquinone"J.Biol.Chem.. 276. 48356-48361 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] O.Adachi: "Membrane-bound quinoprotein D-arabitol dehydrogenase of Gluconobacter suboxydans IFO 3257 : A versatile enzyme for the oxidative fermentation of various ketoses"Biosci.Biotechnol. Biochem.. 65. 2755-2762 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Matsushita: "Quinoproteins : structure, function, and biotechnological applications"Appl.Micriobiol. Biotechnol.. 58. 13-22 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] H.Miyadera: "Quinones in long-lived clk-1 mutants of Caenorhabditis elegans"FEBS Lett.. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Sakamoto: "Synthesis and Electron-transfer Activity o Azido Ubiquinone-2"J.Pesticide Sci.. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 茂木 立志: "細胞呼吸の膜エネルギー装置:起源と分子進化(生体膜のエネルギー装置(吉田賢右, 茂木立志 編)"共立出版. 21 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Uchida,T.: "Resonance Raman studies of oxo intermediates in the reaction of pulsed cytochrome bo with hydrogen peroxide"Biochemistry. 39. 6669-6678 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kobayashi,K.: "Transient formation of ubisemiquinone radical and subsequent electron transfer processes in the Escherichia colicytochrome bo revealed by pulse radiolysis"Biochemistry. 39. 15620-15625 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tsubaki,M.: "Probing molecular structure of dioxygen reduction site of bacterial quinol oxidases through ligand binding to the redox metal centers"J.Inorg.Biochem.. 82. 19-25 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Elias,L.D.: "Functions of amino acid residues in the active site of Escherichia coli PQQ-containing quinoprotein glucose dehydrogenase"J.Biol.Chem.. 275. 7321-7326 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Miyadera,H.: "Altered quinone biosynthesis in the long-kived clk-1 mutants of Caenorhabditis elegans"J.Biol.Chem.. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Matsushita,K.: "Biochemistry and Molecular Biology of vitamin B6 and PQQ-dependent proteins"Birkhauser Verlag AG. 377 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 松下一信: "廣川タンパク質化学 第4巻酵素 4.1 オキシドレダクターゼ"廣川書店. 353 (2000)

    • Related Report
      2000 Annual Research Report

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

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