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リボソーム-RRF複合体の立体構造にもとづく機能解明と創薬への応用

Research Project

Project/Area Number 18054021
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Review Section Biological Sciences
Research InstitutionOsaka University

Principal Investigator

大久保 忠恭  Osaka University, 薬学研究科, 准教授 (90272997)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 2007: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2006: ¥3,000,000 (Direct Cost: ¥3,000,000)
Keywordsリボソーム再生 / 新規抗菌物質 / RRF / NMR / X線結晶構造解析 / ドメイン運動 / リボソーム / EF-G複合体モデル
Research Abstract

リボソームとリボソーム再生因子(RRF)複合体の立体構造をもとに新規抗菌剤の開発を目指して研究を行った。RRFはEF-Gと協同して蛋白質生合成終結後のリボソーム-tRNA-mRNA複合体を解離する。解離後の遊離リボソームは再度開始過程に導かれてリサイクルされる。NMR及びX線結晶構造解析による立体構造の結果、RRFは2つのドメインより構成されること、及びドメインIIがドメイン1に対して角度60度の範囲で揺らぐことを明らかとした。ドメイン1に相当するモデルペプチド(RRF-DI)を合成し相互作用解析を行った結果、 RRFのリボソームとの結合部位はドメイン1であることを明らかにした。RRF-DIとリボソーム50Sサブユニットの複合体の立体構造から構築したRRF、EF-G、70Sリボソーム複合体のモデルから、 RRFの作用機構を明らかにした。さらにRRF-DIとリボソーム50Sサブユニットの結晶構造中の結合部位の立体構造をもとにRRF阻害能を有する分子を設計し評価を行った。RRF単独の溶液構造及び結晶構造をもとにバーチャルスクリーニング法でRRFと結合する低分子分子を探索し、得られた候補化合物数十種類に関して実際に表面プラズモン共鳴法によりRRFとの結合を調べたが、解離定数K_DがサブmMのものしか得られなかった。そこで、RRF-DIとリボソーム50Sサブユニットの結晶構造中でRRFと直接接触しているrRNA部位をテンプレートとする分子を探索した結果、解離定数K_Dが数μMの化合物が得られた。この化合物をもとにRRFとの相互作用様式を考慮して低分子化を試み、同等の解離定数を示す数種類の化合物を得た。

Report

(2 results)
  • 2007 Annual Research Report
  • 2006 Annual Research Report
  • Research Products

    (15 results)

All 2008 2007 2006 Other

All Journal Article (12 results) (of which Peer Reviewed: 7 results) Presentation (1 results) Book (1 results) Remarks (1 results)

  • [Journal Article] NMR structure of an intracellular third loop peptide of human GABA(B) receptor2008

    • Author(s)
      Kikkou, T., et. al.
    • Journal Title

      Biochem Biophys Res Commun. 366

      Pages: 681-684

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Thermal unfolding mechanism of lipocalin-type prostaglandin D synthase2008

    • Author(s)
      Iida, T., et. al.
    • Journal Title

      FEBS J. 275

      Pages: 233-241

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Tautomerism of histidime 64 associated with proton transfer in catalysis of carbonic anhydrase2007

    • Author(s)
      Shimahara, H., et. al.
    • Journal Title

      J. Biol. Chem. 282

      Pages: 9646-9656

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystal structure of Pyrococcus horikoshii PPC protein at 1.60 A resolution2007

    • Author(s)
      Lin, L., et. al.
    • Journal Title

      Proteins 67

      Pages: 505-507

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Crystallization of human nicotinamide phosphoribosyltransferase2007

    • Author(s)
      Takahashi, R., et. al.
    • Journal Title

      Acta Crystallograph Sect. F Struct. Biol. Cryst. Commun. 63

      Pages: 375-377

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] NMR solution structure of lipocalin-type prostaglandin D synthase: Evidence for partial overlapping of catalytic pocket and retinoic acid-binding pocket within the central cavity2007

    • Author(s)
      Shimamoto, S., et. al.
    • Journal Title

      J. Biol. Chem. 282

      Pages: 31373-31379

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Purification, crystallization and preliminary X-ray diffraction study of human ribosomal protein L10 core domain2007

    • Author(s)
      Nishimura, M., et. al.
    • Journal Title

      Acta Crystallograph Sect. F Struct. Biol. Cryst. Commun. 63

      Pages: 950-952

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] NMR structure of a human homologous methuselah gene receptor peptide.2007

    • Author(s)
      Kikkou T., et al.
    • Journal Title

      Biochem Biophys Res. Commun. 352

      Pages: 17-20

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Structure of cytochrome c552 from a moderate thermophilic bacterium, Hydrogenophilus thermoluteolus : comparative study on the thermostability of cytochrome c.2006

    • Author(s)
      Nakamura S, et al.
    • Journal Title

      Biochemistry 45

      Pages: 6115-6123

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Apo- and holo-structures of 3alpha-hydroxysteroid dehydrogenase from Pseudomonas sp. B-0831. Loop-helix transition induced by coenzyme binding.2006

    • Author(s)
      Nakamura S., et al.
    • Journal Title

      J Biol Chem. 281

      Pages: 31876-31884

    • Related Report
      2006 Annual Research Report
  • [Journal Article] NMR structure of an intracellular loop peptide derived from prostaglandin EP3alpha receptor.2006

    • Author(s)
      Kikkou T., et al.
    • Journal Title

      Biochem Biophys Res. Commun. 345

      Pages: 933-937

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Crystallization and preliminary X-ray analysis of the complex of NADH and 3alpha-hydroxysteroid dehydrogenase from Pseudomonas sp. B-0831.2006

    • Author(s)
      Kataoka S., et al.
    • Journal Title

      Acta Crystallograph Sect F Struct Biol Cryst Commun. 62

      Pages: 569-571

    • Related Report
      2006 Annual Research Report
  • [Presentation] リポカリン型プロスタグランジンD合成酵素の溶液構造とリガンド結合様式2007

    • Author(s)
      大久保忠恭
    • Organizer
      特定領域研究「生体超分子の構造形成と機能制御の原子
    • Place of Presentation
      静岡県熱海市ホテルニューア
    • Year and Date
      2007-07-11
    • Related Report
      2007 Annual Research Report
  • [Book] Chromosome Nanoscience and Technology (Ed. Fukui K. and Ushiki T.)2007

    • Author(s)
      Ohkubo, T., et. al.
    • Publisher
      Structure and Interactions of the Imitation SWI-Type Chromatin-Remodeling Complex, ATP-Dependent Chromatin-Assembly Factor
    • Related Report
      2007 Annual Research Report
  • [Remarks]

    • URL

      http://www.phs.osaka-u.ac.jp/homepage/b001/

    • Related Report
      2007 Annual Research Report

URL: 

Published: 2006-04-01   Modified: 2018-03-28  

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