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1988 Fiscal Year Final Research Report Summary

Cytochrome oxidase in thermophilic bacterium PS3 -Crystalization, First and Higher order structure, and H^+ pump activity-

Research Project

Project/Area Number 61480468
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionJichi Medical School

Principal Investigator

SONE Nobuhito  Dept. of Biochem, Jichi Medical School, Associate Prof.., 医学部, 助教授 (20049034)

Co-Investigator(Kenkyū-buntansha) ISHIZUKA Morio  Dept. of Industrial Chem., Tyuo Univ. Associate Prof., 理工学部, 助教授 (50168241)
KAGAWA Yasuo  Dept. of Biochem.,Jichi Medical School, Prof., 医学部, 教授 (30048962)
SAIGO Satoshi  Dept. of Physics, Jichi Medical School, Associate Prof., 医学部, 助教授 (70049071)
Project Period (FY) 1986 – 1988
KeywordsCytochrome oxidase / Thermophilic bacterium / Proton pump / Gene structure / Crystalization of membrane protein / DNA sequence / タン白質一次構造
Research Abstract

1. The genes for subunits of cytochrome oxidase were cloned from a library in EMBL-3 of the thermophilic bacterium PS3 and sequenced. The genes formed an operon in the order of subunits II,I,III and IV. Comparison of the amino acid sequences of each subunit with those from mitochondria and paracoccus denitrificans tells us places of important amino acid residues and a rough image of three-dimensional structure.
2. Crystalization of the enzyme was tried. However, only small crystals seemed to appear, when octylglucoside and heptane triol were added to the mother liquor, which was not suitable for crystalography.
3. Target size of PS3 cytochrome oxidase in respect to both oxidase and proton pump activities were measured. Dimeric structure may be necessary for proton pumping, whereas the monomer can mediate oxidation. The enzyme exist as monomer in several non-ionic detergent solution. Dimeric enzyme may be vormed in the bed of phospholipids.
4. Several methods to measure proton pump activity at the cytochrome oxidase-level have been developed, and applied on several preparations from various bacteria. After consulting several papers on this topics, a review on respiratory complexes in bacteria generating electrochemical proton gradients have been written,
5. Introduction of an expression system will make it possible to confirm putative relationship between structure and function in cytochrome oxidase.

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Sone,N;Kosako,T: EMBO J.5. 1515-1519 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sone,N.: J.Biochem.100. 1465-1470 (1986)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sone,N.;Sekimachi,M.;Kutoh,E.: J.Biochem. 103. 606-610 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sone,N;Yokoi,F;Fu,T.;Ohta,S;Mesto,T.Raitio,M.;Saraste,M.: J.Biol.Chem.103. 606-610 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sone,T.;Takagi,T.: in preparation.

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      「研究成果報告書概要(和文)」より
  • [Publications] Ishizuka,M.;Machida,K.;Shimada,I.;Sone,N.: in preparation.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sone,N. et al.: "Proton pump activity in bacterial cytochrome oxidases in Cytochrome systems(Papa,Chance & Ernster eds)" Plenum press.New York, 743-750 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sone,N. et al.: "Energy-tranducing complexes in bacterial respiratory chains in Subcellular Biochemistry vol.14(Harris & Etemadi eds)" Plenum press,New york, 279-320 (1989)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sone, N.; Kosako, T.: "Evidence for dimer structure of proton-pumping cytochrome coxidase, an analysis by radiation inactivation" EMBO J.5. 1515-1519 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sone, N.: "Measurement of proton pump activity of the thermophilic bacterium PS3 and Nitrobacter agilis at the cytochrome oxidase level using total membranes and heptyl thioglucoside" J. Biochem.100. 1465-1470 (1986)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sone, N.; Sekimachi, M,; Kutoh, E.: "Identification and properties of a quinol oxidase super-complex composed of a bc_1 complex and cytochrome oxidase in the thermophilic bacterium PS3" J. Biol. Chem.262. 15386-15391 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sone, N.; Yokoi, F.; Fu, T.; Ohta, S.; Mesto, T.; Raitio, M.; Saraste, M.: "Nucleotide sequence of the gene coding for cytochrome oxidase subunit I from the thermophilic bacterium PS3" J. Biochem.103. 606-610 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sone, N.; Takagi, T.: "Molecular weights of cytochrome c oxidase and cytochrome bc_1 complex from the thermophilic bacterium PS3"

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sone, N; et al.: Energy-transducing complexes in bacterial respiratory chains as Chapter 7 of Subcellular Biochemistry vol. 14 (Harris and Etemadi eds). 279-320 (1989)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1990-03-20  

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