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Study of mechanics

Research Project

Project/Area Number 12680629
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research Institutionkanazawa University

Principal Investigator

YOKOYAMA Ken  Kanazawa University Department of molecular and collular Biology, 薬学部・分子細胞薬学, 助手 (70271377)

Co-Investigator(Kenkyū-buntansha) OHKUMA Shoji  Kanazawa University Graduate School, Professor, 薬学部・分子細胞薬学, 教授 (10119563)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2000: ¥2,600,000 (Direct Cost: ¥2,600,000)
KeywordsThermus / V-ATPase / T.thermophilus / ATP synthase
Research Abstract

V-type ATPase (V_0V_1) capable of ATP-driven H^+ pumping and of H^+ gradient driven ATP synthesis was isolated from a thermophilic eubacterium, Thermus ther-mophilus. When the enzyme was analyzed by gel electrophoresis in the presence of sodium dodecyl sul-fate, it showed eight polypeptide bands of which four were subunits of V_1. We also isolated the V_0V_1 operon, containing nine genes in the order of atpG-I-L-E-X-F-A-B-D, which encoded proteins with molecular sizes of 13, 43, 10, 20, 35, 11, 64, 53, and 25 kDa, respectively. The last four genes were identified as those for V_0 subunits ; atpA, B, D, and F encoded the A, B, γ, and δ subunits, respectively. The first five genes, atpG-atpX, were identified as genes for the V_0 subunits. The product of atpL, the proteohpid subunit, lacked a 19-amino acid presequence and, unlike V-type ATPases, contained two membrane-spanning domains rather than four. The hydrophobic 43-kDa product of atpl is the smallest member so far found of the eukaryotic 100-kDa subunit family. Its electrophoretic band overlapped with the band of the A subunit. Therefore, all the gene products were found in our purified V_0V_1. We isolated the A_3B_3 subcomplex reconstituted from the isolated subunits and the A_3B_3γ subcomplex from subunit-expressing Escherichia coli. Electron microscopic observation of these subcomplexes revealed that the γ subunit of V_1 filled the central cavity of A_3B_3 and might be central subunit, similar to the γ subunit of F_1ATPase.

Report

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

    (4 results)

All Other

All Publications (4 results)

  • [Publications] Ken Yokoyama その他: "V-type H^+-ATPase/synthase from a Thermophilus"The Journal of Biological Chemistry. 275. 13955-13961 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 13955-13961

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ken Yokoyama, その他: "V-type H^+-ATPase/Synthase from a Thermophilus"THE Journal of Biological chemistry. 275. 13955-13961 (2000)

    • Related Report
      2001 Annual Research Report
  • [Publications] Ken Yokoyama その他5名: "V Type H-ATPase/Synthase from a Thermophilic Eubacterium, Thermus Thermophilus"The Journal of Biological Chemistry. 275. 13955-13961 (2000)

    • Related Report
      2000 Annual Research Report

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

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