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Purification and functional analysis of YhaTU system, a two component K+/H+ antiporter from Bacillus subtilis

Research Project

Project/Area Number 23770156
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Functional biochemistry
Research InstitutionToyo University

Principal Investigator

FUJISAWA Makoto  東洋大学, 食環境科学部, 准教授 (70549641)

Project Period (FY) 2011 – 2013
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2011: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywordsイオンチャネル / トランスポーター / アンチポーター / チャネル / 部位特異的変異 / タンパク質精製 / K+/H+アンチポーター / 枯草菌
Research Abstract

Bacillus subtilis YhaTU system is a K+/H+ antiporter which consists from two components, the hydrophilic ancillary protein YhaT and the membrane-embedded YhaU. The YhaTU system is not only a unique cation proton antiporter but also an interesting enzyme in respect of the evolution of ion antiporter from ion channel. In In this study, purification of the YhaTU system and cloning the genes which have point mutations at the various points of the genes have been tried.
To purify the YhaTU complex, StrepTagII and His6 tags were introduced at N-terminal or C-terminal of the proteins. Fifty six mutations were also introduced into the genes to test the function of each amino acid residues. Now the basis to analyze the detail function of YhaTU has been constructed.

Report

(4 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • 2011 Research-status Report
  • Research Products

    (10 results)

All 2012 2011

All Presentation (10 results)

  • [Presentation] Purification of YhaTU system, a K^+/H^+ a ntiporter from Bacillus subtilis.2012

    • Author(s)
      Kentaro Kato, Yu Tanaami, Makoto Fujisawa
    • Organizer
      The 35th Ann ual Meeting of the Molecular Biology Society of Japan
    • Place of Presentation
      福岡国際会議場、マリンメッセ福岡, 福岡
    • Year and Date
      2012-12-11
    • Related Report
      2013 Final Research Report
  • [Presentation] 枯草菌K+/H+アンチポーターYhaTUシステムの精製 (Purification of YhaTU system, a K+/H+ antiporter from Bacillus subtilis.)2012

    • Author(s)
      加藤健太郎、田名網悠、藤澤誠 (Kentaro Kato, Yu Tanaami, Makoto Fujisawa)
    • Organizer
      第35回日本分子生物学会大会 (The 35th Annual Meeting of the Molecular Biology Society of Japan.)
    • Place of Presentation
      福岡国際会議場・マリンメッセ福岡
    • Related Report
      2012 Research-status Report
  • [Presentation] Detection of YhaTU complex, a Two-Component K^+/H^+ Antiporter from Bacillus subtilis, by Pull-down Assay and Blue Native PAGE.2011

    • Author(s)
      Makoto Fujisawa
    • Organizer
      第9回国際シンポジウムバイオ科学とナノテクノロジーの融合に向けて
    • Place of Presentation
      東洋大学川越キャンパス, 埼玉
    • Year and Date
      2011-12-10
    • Related Report
      2013 Final Research Report
  • [Presentation] プルダウンアッセイとブルーネイティブゲル電気泳動法による枯草菌のK^+/H^+アンチポーターYhaTUシステムの複合体の検出2011

    • Author(s)
      藤澤誠
    • Organizer
      日本農芸化学会関東支部2011年度大会
    • Place of Presentation
      東洋大学板倉キャンパス, 群馬
    • Year and Date
      2011-10-15
    • Related Report
      2013 Final Research Report
  • [Presentation] Detection of YhaTU complex, a Two-Component K^+/H^+ Antiporter from Bacillus subtilis, by Pull-down Assay and Blue Native PAGE.2011

    • Author(s)
      Makoto Fujisawa
    • Organizer
      International Union of Microbiological Societies 2011 Congress
    • Place of Presentation
      札幌コンベンションセンター, 北海道
    • Related Report
      2013 Final Research Report
  • [Presentation] Detection of interaction between subunits of a two-component K^+/H^+.2011

    • Author(s)
      Makoto Fujisawa
    • Organizer
      Gordon Research Conference–Mechanisms of Membrane
    • Place of Presentation
      University of New England, 米国メイン州
    • Related Report
      2013 Final Research Report
  • [Presentation] Detection of YhaTU complex, a Two-Component K+/H+ Antiporter from Bacillus subtilis, by Pull-down Assay and Blue Native PAGE.2011

    • Author(s)
      Makoto Fujisawa
    • Organizer
      9th International Symposium on Bioscience and Nanotechnology
    • Place of Presentation
      Toyo University, Kawagoe, Saitama, Japan
    • Related Report
      2011 Research-status Report
  • [Presentation] プルダウンアッセイとブルーネイティブゲル電気泳動法による枯草菌のK+/H+アンチポーターYhaTUシステムの複合体の検出2011

    • Author(s)
      藤澤 誠
    • Organizer
      日本農芸化学会関東支部2011年度大会
    • Place of Presentation
      東洋大学 板倉キャンパス(群馬県)
    • Related Report
      2011 Research-status Report
  • [Presentation] Detection of YhaTU complex, a Two-Component K+/H+ Antiporter from Bacillus subtilis, by Pull-down Assay and Blue Native PAGE.2011

    • Author(s)
      Makoto Fujisawa
    • Organizer
      International Union of Microbiological Societies 2011 Congress
    • Place of Presentation
      Sapporo Convention Center,Hokkaido,Japan
    • Related Report
      2011 Research-status Report
  • [Presentation] Detection of interaction between subunits of a two-component K+/H+ antiporter, YhaTU, from Bacillus subtilis.2011

    • Author(s)
      Makoto Fujisawa
    • Organizer
      Gordon Research Conference - Mechanisms of Membrane Transport
    • Place of Presentation
      University of New England, Biddeford, ME, USA
    • Related Report
      2011 Research-status Report

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Published: 2011-08-05   Modified: 2019-07-29  

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