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チトクロム酸化酵素結晶の高精度高分解能X線回折強度データ収集システムの開発

Research Project

Project/Area Number 18770088
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Structural biochemistry
Research InstitutionOsaka University

Principal Investigator

山下 栄樹  Osaka University, 蛋白質研究所, 助教 (00294132)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2007: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2006: ¥1,600,000 (Direct Cost: ¥1,600,000)
Keywordsチトクロム酸化酵素 / 膜蛋白質複合体 / 結晶構造解析 / 放射光
Research Abstract

チトクロム酸化酵素は呼吸鎖の末端に位置し,チトクロムcから電子を受け取り分子状酸素を水にまで還元するとともに、そのとき得られる自由エネルギーによって、プロトンを膜の内側から外側へと能動輸送を行う膜貫通型膜蛋白質複合体である。このチトクロム酸化酵素のプロトン能動輸送機構を解明するためには、酵素内の水素原子の座標を決定する必要がある。このためには、高分解能高精度のX線回折強度データを収集し、精密な構造を得る必要がある。本研究では、放射光の大強度で平行性の良いX線を利用し、高分解能の回折斑点が得られるチトクロム酸化酵素結晶を効率良く選択できるシステムを構築するとともに、高分解能高精度のX線回折強度データを収集するための測定法の開発を行なった。
平成18年度に本研究で開発しSPring-8生体超分子複合体ビームライン(BL44XU)に導入した1枚のX線回折像から回折斑点のS/Nを計算することにより分解能を見積るソフトウェアを用い、チトクロム酸化酵素の回折撃度測定において入射X線の条件や測定方法の検討を行った結果、入射X線の密度を高くすることが最も効果的で、1.8Å以上の回折強度測定が可能になった。

Report

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

    (22 results)

All 2008 2007 2006

All Journal Article (11 results) (of which Peer Reviewed: 6 results) Presentation (11 results)

  • [Journal Article] Structures and physiological roles of 13 integral lipids of bovine heart cytochrome c oxidase2007

    • Author(s)
      Shinzawa-Itoh K.
    • Journal Title

      EMBO J. 26

      Pages: 1713-1725

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The.Crystal structure of a Virus-like Particle from the Hyperthermophilic Archaeon Pyrococcus furiosus Provides Insight into the Evolution of Viruses.2007

    • Author(s)
      Akita F.
    • Journal Title

      J Mol.Biol. 368

      Pages: 1469-1483

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A histidine residue acting as a controlling site for dioxygen reduction and proton pumping by cytochrome c oxidase2007

    • Author(s)
      Muramoto K.
    • Journal Title

      Proc Natl Acad Sci U S A 104

      Pages: 7881-7886

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Structure of the cytochrome b6f complex: quinone analogue inhibi tors as li sands of heme cn2007

    • Author(s)
      Yamashita E.
    • Journal Title

      J Mol. Biol. 370

      Pages: 39-52

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] structure of the Complex of the Colicin E2 R-domain and lts BtuB Receptor: THE OUTER MEMBRANE COLICIN TRANSLOCON2007

    • Author(s)
      Sharma 0.
    • Journal Title

      J. Biol. Chem. 282

      Pages: 23163-23170

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Ultrafast Optical Pump-Probe studies of the Cytochrome b(6)f Complex in Solution and Crystalline states.2007

    • Author(s)
      Dashdorj N.
    • Journal Title

      J.Phys. Cbem. B 111

      Pages: 14405-14410

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The asymmetric IscA homodimer with an exposed [2Fe-2S]cluster suggests the structural basis of the Fe-S cluster biosynthetic scaffold.2006

    • Author(s)
      Morimoto K., Yamashita E., Kondou Y., Lee S.J., Arisaka F., Tsukihara T., Nakai M.
    • Journal Title

      JournaL of molecular Biology 360

      Pages: 117-132

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Crystal tructures of the catalytic domain of human stromelysin-1(MMP-3) and collagenase-3 (MMP-13) with a hydroxamic acid inhibitor SM-254532006

    • Author(s)
      Kohno T., Hochigai H., Yamashita E., Tsukihara T., Kanaoka M.
    • Journal Title

      Biochemical and Biophysical Research Communications 344

      Pages: 315-322

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Crystal structures of a multidrug transporter reveal a functionally rotating mechanism2006

    • Author(s)
      Murakami S., Nakashima R., Yamashita E., Matsumoto T., Yamaguchi A.
    • Journal Title

      Nature 443

      Pages: 173-179

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Crystal Structure of DsbB-DsbA Complex Revealing a Cysteine Relocation Mechanism2006

    • Author(s)
      Inaba K., Murakami S., Suzuki M., Nakagawa A., Yamashita E., Okada K., Ito K.
    • Journal Title

      Cell 127

      Pages: 789-801

    • Related Report
      2006 Annual Research Report
  • [Journal Article] In Meso Structure of the Cobalamin Transporter, BtuB, at 1.95 A Resolution2006

    • Author(s)
      Cherezov V., Yamashita E., Liu W., Zhalnina M., Cramer W.A., Caffrey M.
    • Journal Title

      JournaL of molecular Biology 364

      Pages: 716-734

    • Related Report
      2006 Annual Research Report
  • [Presentation] SPring-8生体超分子複合体構造解析ビームライン(BL44XU)2008

    • Author(s)
      吉村政人
    • Organizer
      第21回日本放射光学会年会・放射光科学合同シンポジウム
    • Place of Presentation
      立命館大学びわこ草津キャンパス
    • Year and Date
      2008-01-14
    • Related Report
      2007 Annual Research Report
  • [Presentation] SPring-8生体超分子複合体構造解析ビームライン(BL44XU)の現状2008

    • Author(s)
      山下栄樹
    • Organizer
      第21回日本放射光学会年会・放射光科学合同シンポジウム
    • Place of Presentation
      立命館大学びわこ草津キャンパス
    • Year and Date
      2008-01-13
    • Related Report
      2007 Annual Research Report
  • [Presentation] ウシ心筋チトクロム酸化酵素の反応中間体のX線構造解析2007

    • Author(s)
      鍋倉 弘行
    • Organizer
      日本生物物理学会第45回年会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-23
    • Related Report
      2007 Annual Research Report
  • [Presentation] ウシ心筋チトクロム酸化酵素のZn阻害部位のX線構造解析2007

    • Author(s)
      村本 和優
    • Organizer
      日本生物物理学会第45回年会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-23
    • Related Report
      2007 Annual Research Report
  • [Presentation] 部分還元型ウシ心筋チトクロム酸化酵素のX線構造解析2007

    • Author(s)
      谷口 槙
    • Organizer
      日本生物物理学会第45回年会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-23
    • Related Report
      2007 Annual Research Report
  • [Presentation] シアン化物結合型チトクロム酸化酵素のX線結晶構造解析2007

    • Author(s)
      望月 正雄
    • Organizer
      日本生物物理学会第45回年会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-23
    • Related Report
      2007 Annual Research Report
  • [Presentation] 酸化型ウシ心筋チトクロム酸化酵素のX線構造2007

    • Author(s)
      青山 浩
    • Organizer
      日本生物物理学会第45回年会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-23
    • Related Report
      2007 Annual Research Report
  • [Presentation] structure of the Cytochrome b_6f Compex: Quinone Analogue Inhibitors as Ligands of Heme c_n2007

    • Author(s)
      Yamashita E.
    • Organizer
      The 8th Conference of the Asian Crystallographic Association
    • Place of Presentation
      Howard International House Taipei, Taiwan R. O. C.
    • Year and Date
      2007-09-06
    • Related Report
      2007 Annual Research Report
  • [Presentation] The crystal structure of a virus-like particle from hyperthermophillic arcaheon pyrococcus furiosus provide insight into the evolution of viruses2007

    • Author(s)
      Akita F.
    • Organizer
      The 8th Conference of the Asian Crystallographic Association
    • Place of Presentation
      Howard International House Taipei, Taiwan R.0.C.
    • Year and Date
      2007-09-05
    • Related Report
      2007 Annual Research Report
  • [Presentation] 超好熱古細菌Pyrococcus furiosus由来ウィルス様粒子の自己組織化及び耐熱機構について2007

    • Author(s)
      秋田総理
    • Organizer
      第7回日本蛋白質科学会年会
    • Place of Presentation
      仙台国際センター
    • Year and Date
      2007-05-26
    • Related Report
      2007 Annual Research Report
  • [Presentation] ラット肝臓由来vaultのX線結晶構造解析2007

    • Author(s)
      田中秀明
    • Organizer
      日本結晶学会2007年度年会
    • Place of Presentation
      東京工業大学・大岡山キャンパス
    • Related Report
      2007 Annual Research Report

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Published: 2006-04-01   Modified: 2016-04-21  

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