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DEVELOPMENT OF TERMINAL OXIDASE-SPECIFIC BACTERIOCIDES AND BIOCHEMICAL ANALYSIS OF THE SUBSTRATE OXIDATION MECHANISM

Research Project

Project/Area Number 07558221
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field Functional biochemistry
Research InstitutionTHE UNIVERSITY OF TOKYO

Principal Investigator

MOGI Tatsushi  THE UNIVERSITY OF TOKYO,GRADUATE SCHOOL OF SCIENCE,RESEARCH ASST., 大学院・理学系研究科, 助手 (90219965)

Co-Investigator(Kenkyū-buntansha) NAGAMURA Toshihiko  YUNISOKU CO., DEVELOP.RESEARCH INST,DIRECTOR, 科学機器開発研究所, 所長
MIYOSHI Hideto  THE UNIVERSITY OF TOKYO,FAC.OF AGRICULTURE,KYOTO UNIV., ASST PROFESSOR, 農学部, 助教授 (20190829)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1996: ¥2,500,000 (Direct Cost: ¥2,500,000)
KeywordsAnti-bacteriocide / Terminal oxidase / Quinol oxidase / Quinone analogue / Escherichia coli / Substrate oxidation site / Substituted phenol / Reaction mechanism / 基質酸化機構 / 耐性変異 / 置換フェノール類 / ユビキノン / 活性評価システム / カプサイシン
Research Abstract

The aims of this project are to develop new bacteriocides targetted to quinol oxidases of the aerobic respiratory chain and to elucidate biochemically the molecular mechanism of the substrate oxidation. Results obtained are follows ;
1. Based on structure-function studies on ubiquinone analogues, we found that the substrate oxidation sites (Q_I) of the Escherichia coli cytochromes bo and bd, which are not structurally related, have steric constraints against the 2-methoxy and 5-methyl groups. In addition, the 6-isoprenyl group increases the binding affinity of cytochrome bo.
2. Substrate specificity of the high affinity quinone binding site (Q_H) of cytochrome bo was different from that of the Q_L site. Reconstitution of the Q_H site with substituted phenols, flow-flash studies of the reduced Q_H-free enzyme with dioxygen and redox titrations indicated that the Q_H site mediates electron transfer from the Q_L site to low-spin heme b.
3. Screening of antimycin A,rorenone and capsaicin derivatives revealed that some antimycin A derivatives can inhibit quinol oxidases strongly.
4. Among new quinone analogues identified in this study, three compounds were shown to inhibit the aerobic growth of E.coli and can be used as a lead compound for design of terminal oxidase-specific bacteriocides. Then, we isolated and characterized the quinone analogue-resistant mutants of cytochrome bo biochemically and molecular biologically. Results indicate that two quinone/quinol binding sites in cytochrome bo function cooperatively to catalyze the quinol oxidation and electron transfer from the Q_L to heme b in subunit I.
5. Molecular biological and chemical cross-linking studies indicate that the structure of cytochrome bo is similar to that of cytohcrome c oxidase from Paracoccus denitrificans. To determine an atomic structure of cytochrome bo, the examination of the crystallization conditions is in progress.

Report

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

    (65 results)

All Other

All Publications (65 results)

  • [Publications] M.Sato-Watanabe: "Stabilization of a semiquinone radical at the high affinity quinone binding site of the Escherichia coli bo-type ubiquinol oxidase." FEBS Lett.374. 265-269 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Y.Orii: "Facilitated intramolecular electron transfer in the Escherichia coli bo-type ubiquinol oxidase requires chloride." Biochemistry. 34. 1127-1132 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Mogi: "Cu_B promotes both binding and reduction of dioxygen at the heme-copper binuclear center in the Escherichia coli bo-type ubiquinol oxidase." FEBS Lett.370. 259-263 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] S.Hirota: "Resonance Raman study on axial ligands of heme irons in cytochrome bd-type ubiquinol oxidase from Escherichia coli." Biospectroscopy. 1. 305-311 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Tsubaki: "Cyanide-binding site of bd-type ubiquinol oxidase from Escherichia coli." J.Biol.Chem.270. 28565-28569 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Saitoh: "Comparison of the inhibitory action of synthetic capsicin analogues with various NADH-ubiquinone oxidoreductases." Biochim.Biophys.Acta. 1273. 21-30 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Denda: "Characterization of chimeric heme-copper respiratory oxidases using subunits I of Escherichia coli cytochrome bo and Halobacterium salinarium cytochrome ca." Biochem.Biophys.Res.Commun.217. 428-436 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 茂木立志: "大腸菌チトクロムbo型ユビキノール酸化酵素の構造と分子生理学" 生化学. 67. 93-107 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Miyoshi: "A model of antimycin A binding based on structure-activity studies of synthetic antimycin A analogues." Biochim.Biophys.Acta. 1229. 149-154 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 三芳秀人: "阻害剤から見た呼吸鎖電子伝達機構" 化学と生物. 33. 151-158 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Tsubaki: "FT-IR and EPR studies on cyanide-bindings to the heme-copper binuclear center of cytochrome bo-type ubiquinol oxidase from Escherichia coli : Release of Cu_B-cyano complex in the partially reduced states." J.Biol.Chem.271. 4017-4022 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Hori: "EPR and NO complex of bd-type ubiquinol oxidase from Escherichia coli. The proximal ligand of heme d is a nitrogenous amino acid residue." J.Biol.Chem.271. 9254-9258 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Saiki: "Probing a role of subunit IV of the Escherichia coli bo-type ubiquinol oxidase by deletion and cross-linking analyses." J.Biol.Chem.271. 15336-15340 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Sakamoto: "Probing substrate binding site of the Escherichia coli quinol oxidases using synthetic ubiquinol analogues based upon their electron-donating efficiency." J.Biol.Chem.271. 29897-29902 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Sakamoto: "Comparison of the structural features of ubiquinone reduction sites between glucose dehydrogenase in Escherichia coli and bovine heart mitochondrial complex I." Eur.J.Biochem.237. 128-135 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] C.Lim: "Phytotoxic compounds cochlioquinones are inhibitors of mitochondrial NADH-ubiquinone reductase." J.Pesticide Science. 21. 213-215 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Miyoshi: "Uncoupling mechanisms of glycoside antibiotic aculeximycin in isolated rat-liver mitochondria." J.Biochem.119. 274-280 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] A.Yamashita: "Direct interaction between mitochondrial succinate-ubiquinone and ubiquinol-cytochrome c oxidoreductase probed by inhibitor sensitivity." J.Biochem.120. 377-384 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Satoh: "Comparison of the inhibitory action of synthetic capsaicin analogues with various NADH-ubiquinone oxidoreductases." Biochim.Biophys.Acta. 1273. 21-30 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Ueno: "Structural factors of rotenone required for inhibition of various NADH-ubiquinone oxidoreductases." Biochim.Biophys.Acta. 1276. 195-202 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] V.Yankovskaya: "Inhibitor probes of the quinone binding sites of mammalian complex II and Escherichia coli fumarate reductase." J.Biol.Chem.271. 21020-21024 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Mogi: "Molecular structure and physiology of cytochrome bo-type ubiquinol oxidase from Escherichia coli" Seikagaku. 67. 93-107 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Miyoshi: "Inhibitors and respiratory electron-transport mechanism." Kagaku to Seibutsu. 33. 151-158 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Mogi: "Cu_B promotes both binding and reduction of dioxygen at the heme-copper binuclear center in the Escherichia coli bo-type ubiquinol oxidase." FEBS Lett.370. 259-263 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Sato-Watanabe: "Stabilization of a semiquinone radical at the high affinity quinone binding site of the Escherichia coli bo-type ubiquinol oxidase." FEBS Lett.374. 265-269 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] S.Hirota: "Resonance Raman study on axial ligands of heme irons in cytochrome bd-type ubiquinol oxidase from Escherichia coli." Biospectroscopy. 1. 305-311 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Tsubaki: "Cyanide-binding site of bd-type ubiquinol oxidase from Escherichia coli." J.Biol. Chem.270. 28565-28569 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Denda: "Characterization of chimeric heme-copper respiratory oxidases using subunit of Escherichia coli cytochrome bo and Halobacterium salinarium cytochrome omega_3." Biochem. Biophys. Res. Commun.217. 428-436 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Saitoh: "Comparison of the inhibitory action of synthetic capsicin analogues with various NADH-ubiquinone oxidoreductases." Biochim. Biophys. Acta. 1273. 21-30 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Miyoshi: "A model of antimycin A binding based on structure-activity studies of synthetic antimycin A analogues." Biochim. Biophys. Acta. 1229. 149-154 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Tsubaki: "FT-IR and EPR studies on cyanide-bindings to the heme-copper binuclear center of cytochrome bo-type ubiquinol oxidase from Escherichia coli : Release of Cu_B-cyano complex in the partially reduced states." J.Biol. Chem.271. 4107-4022 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Hori: "EPR and NO complex of bd-type ubiquinol oxidase from Escherichia coli. The proximal ligand of heme d is a nitrogenous amino acid residue." J.Biol. Chem.271. 9254-9258 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Saiki: "Probing a role of subunit IV of the Escherichia coli bo-type ubiquinol oxidase by deletion and cross-linking analyzes." J.Biol. Chem.271. 15336-15340 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Sakamoto: "Probing substrate binding site of the Escherichia coli quinol oxidases using synthetic ubiquinol analogues based upon their electron-donating efficiency." J.Biol. Chem.271. 29897-29902 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] K.Sakamoto: "Comparison of the structural features of ubiquinone reduction sites between glucose dehydrotgenase in Escherichia coli and bovine heart mitochondrial complex I." Eur. J.Biochem.237. 128-135 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] C.-H.Lim: "Phytotoxic compounds cochlioquinones are inhibitors of mitochondrial NADH-ubiquinone reductase." J.Pesticide Science. 21. 213-215 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Miyoshi: "Uncoupling mechanisms of glycoside antibiotic aculeximycin in isolated rat-liver mitochondria." J.Biochem.119. 274-280 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] A.Yamashita: "direct interaction between mitochondrial succinate-ubiquinone and ubiquinol-cytochrome c oxidoreductase probed by inhibitor sensitivity." J.Biochem.120. 377-384 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Satoh: "Comparison of the inhibitory action of synthetic capsaicin analogues with various NADH-ubiquinone oxidoreductases." Biochim. Biophys. Acta. 1273. 21-30 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Ueno: "Structural factors of rotenone required for inhibition of various NADH-ubiquinone oxidoreductases." Biochim. Biophys. Acta. 1276. 195-202 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] V.Yankovskaya: "Inhibitor probes of the quinone binding sites of mammalian complex II and Escherichia coli fumarate reductase." J.Biol. Chem.271. 21020-21024 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Sato-Watanabe: "Cooperation of two quinone binding sites in the oxidation of substrates by cytochrome bo." Oxygen Homeostasis and ts Dynamics (ed. Y.Ishimura). Springer-Verlag, Tokyo.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Tsbaki,T.Mogi,H.Hori,M.Sato-Watanabe,and Y.Anraku: "FT-IR and EPR studies on cyanide-bindings to the heme-copper binuclear center of cytochrome bo-type ubiquinol oxidase from Escherichia coli:Release of CuB-cyano complex in the partially reduced states." J.Biol.Chem. 271. 4017-4022 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Hori,M.Tsubaki.T.Mogi,and Y.Anraku: "EPR and NO complex of bd-type ubiquinol oxidase from Escherichia coli. The proximal ligand of heme d is a nitrogeneous amino acid residue." J.Biol.Chem. 271. 9254-9258 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Saiki,T.Mogi,and Y.Anraku: "Probingg a role of subunit IV of the Escherichia coli bo-type ubiquinol oxidase by deletion and cross-linking analyses." J.Biol.Chem. 271. 15336-15340 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Sakamoto,H.Miyoshi,K.Takegami,T.Mogi,Y.Anraku,and H.Iwamura: "Probing substrate bindings site of the Eschericia coli quinol oxidases using synthetic ubiquinol analogues based upon their electro-donating efficiency." J.Biol.Chem. 271. 29897-29902 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Sakamoto,H.Miyoshi,K.Matsushita,M.Nakagawa,I.Ikeda,M.Ohshima,O.Adachi,T.Akagi,and H.Iwamura: "Comparison of the structural features of ubiquinone reduction sites between glucose dehydrogenase in Escherichia coli and bovine heart mitochondrial complex I." Eur.J.Biochem.237. 128-135 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] C.Lim,H.,H.Ueno,H.Miyoshi,H.Miyagawa,H.Iwamura,and T.Ueno: "Phytotoxic compounds cochlioquinones are inhibitors of mitochondrial NADH-ubiquinone reductase." J.Pesticide Science. 21. 213-215 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Miyoshi,M.Tamaki,H.Murata,T.Ikemoto,T.Shibuya,K.I.Harada,M.Suzuki,and H.Iwamura: "Uncoupling mechanisms of glycoside antibiotic aculeximycin in isolated rat-liver mitochondria." J.Biochem.119. 274-280 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] A.Yamashita,H.Miyoshi,T.Hatano,and H.Iwamura: "Direct interaction between mitochondrial succinate-ubiquinone and ubiquinol-cytochrome c oxidoreductase probed by inhibitor sensitivity." J.Biochem.120. 377-384 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Satoh,H.Miyoshi,K.Sakamoto and H.Iwamura: "Comparison of the inhibitory action of synthetic capsaicin analogues with various NADH-ubiquinone oxidoreductases." Biochim.Biophys.Acta. 1273. 21-30 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] H.Ueno,H.Miyoshi,M.Inoue,Y.Niidome and H.Iwamura: "Structural factors of rotenone required for inhibition of various NADH-ubiquinone oxidoreductases." Biochim.Biophys.Acta. 1276. 195-202 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] V.Yankovskaya,V.,S.O.Salbin,R.R.Ramsay,T.P.Singer and H.Miyoshi: "Inhibitor probes of the quinone binding sites of mammalian complex II and Escherichia coli fumarate reductase." J.Biol.Chem. 271. 21020-21024 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] M. Sato-Watanabe: "Stabilization of a semiquinone radical at the high affinity quinone binding site of the Escherichia coil bo-type ubiquinol oxidase." FEBS Lett.374. 265-269 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Y. Orii: "Facilitated intramolecular electron transfer in the Escherichia coli bo-type ubiquinol oxidase requires chloride" Biochemistry. 34. 1127-1132 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T. Mogi: "CuB promotes both binding and reduction of dioxygen at the heme-copper binuclear center in the Escherichia coli bo-type ubiquinol oxidase." FEBS Lett.370. 259-263 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] S. Hirota: "Resonance Raman study on axial ligands of heme irons in cytochrome bd-type ubiquinol oxidase from Escherichia coli" Biospectroscopy. 1. 305-311 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M. Tsubaki: "Cyanide-binding site of bd-type ubiquinol oxidase from Escherichia coli." J. Biol. Chem.270. 28565-28569 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M. Tsubaki: "FT-IR and EPR studies on cyanide-bindings to the heme-copper binuclear center of cytochrome bo-type ubiquinol oxidase from Escherichia coli:Release of CuB-cyano complex in the partially reduced states." J. Biol. Chem.

    • Related Report
      1995 Annual Research Report
  • [Publications] K. Denda: "Characterization of chimeric heme-copper respiratory oxidases using subunits I of Escherichia coli cytochrome bo and Halobacterium salinarium cytochrome aa_3." Biochem. Biophys. Res. Commun.217. 428-436 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 茂木立志: "大腸菌チトクロムbo型ユビキノール酸化酵素の構造と分子生理学" 生化学. 67. 93-107 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K. Sakamoto: "Comparison of the structural features of ubiquinone reduction sites between glucose dehydrogenase in Escherichia coli and bovine heart mitochondrial complex I." Eur. J. Biochem. (印刷中). (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] T. Saitoh: "Comparison of the inhibitory action of synthetic capsicin analogues with various NADH-ubiquinone oxidoreductases." Biochim. Biophys. Acta. (印刷中). (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] H. Miyoshi: "A model of antimycin A binding based on structure-activity studies of synthetic antimycin A analogues." Biochim. Biophys. Acta. 1229. 149-154 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 三芳秀人: "阻害剤から見た呼吸鎖電子伝達機構" 化学と生物. 33. 151-158 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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