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Structural analyses of MAPEG family membrane proteins using NMR

Research Project

Project/Area Number 26460038
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Physical pharmacy
Research InstitutionYokohama City University

Principal Investigator

Sakakura Masayoshi  横浜市立大学, 生命医科学研究科, 助教 (20334336)

Co-Investigator(Kenkyū-buntansha) 高橋 栄夫  横浜市立大学, 生命医科学研究科, 教授 (60265717)
Research Collaborator FUJII Moe  横浜市立大学, 大学院生命医科学研究科, 技術補助員
FUSHIMI Taketoshi  横浜市立大学, 大学院生命医科学研究科, 大学院生
KOIKE Kenichiro  横浜市立大学, 大学院生命医科学研究科, 大学院生
SUZUKI Rika  横浜市立大学, 大学院生命医科学研究科, 大学院生
Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywords構造生物学
Outline of Final Research Achievements

Eicosanoids are chemical mediators synthesized from arachidonic acid. Three tetra-span trimeric membrane proteins, FLAP, LTC4S, and mPGES1, are (co)-enzymes involved in this arachidonic acid metabolism, and are thought to be drug targets. To understand molecular mechanisms for these proteins to function, we performed structural analyses of these membrane proteins using NMR. We expressed these proteins by using the yeast Pichia pastoris expression system, and found the yield of FLAP is more than 30 times higher than that of the E. coli system. We then reconstituted these proteins in detergent micelles and/or nanolipoprotein particles. The NMR interaction study of FLAP and its inhibitor MK-591 revealed that amino acids located more than 10 angstrom apart from the ligand in the crystal structure experienced chemical shift change by the addition of the inhibitor, which suggests the MK-591 binding induces conformational changes around the transmembrane helices of FLAP.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (9 results)

All 2017 2016 2015 2014

All Presentation (9 results) (of which Int'l Joint Research: 1 results)

  • [Presentation] NMRを用いたヒト膜タンパク質FLAPの物理化学的性状解析2017

    • Author(s)
      森雅樹、坂倉正義、鈴木里佳、伏見威俊、高橋栄夫
    • Organizer
      日本薬学会第137年会
    • Place of Presentation
      仙台国際センター(仙台市青葉区)
    • Year and Date
      2017-03-24
    • Related Report
      2016 Annual Research Report
  • [Presentation] 酵母 Pichia pastoris 発現系を用いて調製した安定同位体標識ヒト膜タンパク質のNMR解析2016

    • Author(s)
      坂倉正義、鈴木里佳、森雅樹、藤井萌、高橋栄夫
    • Organizer
      第55回NMR討論会
    • Place of Presentation
      広島国際会議場(広島市中区)
    • Year and Date
      2016-11-16
    • Related Report
      2016 Annual Research Report
  • [Presentation] Methyl-selective isotope labeling strategy using the yeast Pichia pastoris2016

    • Author(s)
      Rika Suzuki, Masayoshi Sakakura, Moe Fujii, Masaki Mori, and Hideo Takahashi
    • Organizer
      XXVIIth International Conference on Magnetic Resonance in Biological Systems
    • Place of Presentation
      京都国際会館(京都市左京区)
    • Year and Date
      2016-08-21
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] NMRを用いた脂質二重膜中におけるヒト膜タンパク質C99の解析2016

    • Author(s)
      真嶋健大、坂倉正義、藤井萌、三尾和弘、高橋栄夫
    • Organizer
      日本薬学会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2016-03-26
    • Related Report
      2015 Research-status Report
  • [Presentation] NMRを用いた脂質二重膜中におけるヒト膜タンパク質の解析2015

    • Author(s)
      坂倉正義、真嶋健大、藤井萌、太田修平、小池賢一郎、高橋栄夫
    • Organizer
      日本分子生物学会
    • Place of Presentation
      神戸ポートアイランド
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] NMR解析を指向した酵母発現系による安定同位体標識ヒト膜タンパク質の調製2015

    • Author(s)
      鈴木里佳、坂倉正義、太田修平、伏見威俊、高橋栄夫
    • Organizer
      日本分子生物学会
    • Place of Presentation
      神戸ポートアイランド
    • Year and Date
      2015-12-01
    • Related Report
      2015 Research-status Report
  • [Presentation] NMR解析を指向した酵母発現系による安定同位体標識ヒト膜タンパク質試料の調製2015

    • Author(s)
      鈴木里佳、坂倉正義、伏見威俊、小池賢一郎、高橋栄夫
    • Organizer
      日本薬学会第135年会
    • Place of Presentation
      神戸サンボーホール(兵庫県神戸市)
    • Year and Date
      2015-03-25 – 2015-03-28
    • Related Report
      2014 Research-status Report
  • [Presentation] NMR構造解析に向けたヒト膜タンパク質の酵母発現系の構築2014

    • Author(s)
      坂倉正義、伏見威俊、小池賢一郎、鈴木里佳、山田有紗、高橋栄夫
    • Organizer
      第37回日本分子生物学会年回
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] NMR解析を指向したヒト膜タンパク質の酵母発現系の構築2014

    • Author(s)
      坂倉正義、伏見威俊、小池賢一郎、高橋栄夫
    • Organizer
      第14回日本蛋白質科学会年会
    • Place of Presentation
      横浜産貿ホール マリネリア(神奈川県横浜市)
    • Year and Date
      2014-06-25 – 2014-06-27
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2018-03-22  

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