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Structural analysis of type III protein export apparatus by electron cryomicroscopy

Research Project

Project/Area Number 18K14639
Research Category

Grant-in-Aid for Early-Career Scientists

Allocation TypeMulti-year Fund
Review Section Basic Section 43020:Structural biochemistry-related
Research InstitutionOsaka University

Principal Investigator

Kawamoto Akihiro  大阪大学, 蛋白質研究所, 助教 (90631523)

Project Period (FY) 2018-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywordsクライオ電子顕微鏡 / タンパク質輸送 / 膜タンパク質 / Ⅲ型分泌系 / 蛋白質輸送 / 構造生物学 / 細菌 / III型分泌系 / 病原性細菌
Outline of Final Research Achievements

In this study, we aimed to elucidate the mechanism of protein secretion in the type III secretion system by using cryo-electron microscopy to reveal the overall structure of the system at high resolution.
We focused on the bacterial flagellar type III protein secretion system, which is classified into the same type III secretion system, and analyzed the structure of the constitutive membrane protein, revealing that it form symmetries different from those predicted by previous biochemical experiments. In addition, the structure is similar to the previously reported ion channels suggested that the function were also similar.

Academic Significance and Societal Importance of the Research Achievements

多剤耐性菌の出現により治療困難な感染症が増え、新たな視点からの感染症対策が急務となっている。本研究で着目しているⅢ型分泌系の蛋白質分泌機構を分子・原子レベルで解明し、これらの働きを不活化することができれば、細菌を殺さず病原性のみを破壊する新しいタイプの感染症治療薬の開発につながることが期待される。また、すでに報告されているイオンチャネルの構造との類似性が示されたことから、これまで提唱されていた分泌機構とは異なる、新たな蛋白質の分泌機構の解明につながることが期待できる。

Report

(3 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • Research Products

    (21 results)

All 2020 2019 2018 Other

All Int'l Joint Research (4 results) Journal Article (9 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 9 results,  Open Access: 7 results) Presentation (7 results) (of which Invited: 2 results) Remarks (1 results)

  • [Int'l Joint Research] EMBL Australia/School of Medical Sciences/UNSW Sydney(オーストラリア)

    • Related Report
      2019 Annual Research Report
  • [Int'l Joint Research] Physical& Theoretical Chemistry/Department of Chemistry/University of Oxford(英国)

    • Related Report
      2019 Annual Research Report
  • [Int'l Joint Research] University of New South Wales(オーストラリア)

    • Related Report
      2018 Research-status Report
  • [Int'l Joint Research] Oxford University(英国)

    • Related Report
      2018 Research-status Report
  • [Journal Article] Cryo-EM structures reveal translocational unfolding in the clostridial binary iota toxin complex2020

    • Author(s)
      Yamada Tomohito、Yoshida Toru、Kawamoto Akihiro、Mitsuoka Kaoru、Iwasaki Kenji、Tsuge Hideaki
    • Journal Title

      Nature Structural & Molecular Biology

      Volume: 27 Issue: 3 Pages: 288-296

    • DOI

      10.1038/s41594-020-0388-6

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] In Vitro Autonomous Construction of the Flagellar Axial Structure in Inverted Membrane Vesicles.2020

    • Author(s)
      Terashima H, Tatsumi C, Kawamoto A, Namba K, Minamino T, Imada K.
    • Journal Title

      Biomolecules

      Volume: 10 Issue: 1 Pages: 126-126

    • DOI

      10.3390/biom10010126

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Refined mechanism of Mycoplasma mobile gliding based on structure, ATPase activity, and sialic acid binding of machinery2019

    • Author(s)
      Nishikawa MS, Nakane D, Toyonaga T, Kawamoto A, Kato T, Namba K, Miyata M
    • Journal Title

      mBio

      Volume: 10 Issue: 6 Pages: 1-12

    • DOI

      10.1128/mbio.02846-19

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Design and synthesis of pleated DNA origami nanotubes with adjustable diameters2019

    • Author(s)
      Berengut Jonathan F、Berengut Julian C、Doye Jonathan P K、Preern Domen、Kawamoto Akihiro、Ruan Juanfang、Wainwright Madeleine J、Lee Lawrence K
    • Journal Title

      Nucleic Acids Research

      Volume: 47 Pages: 11963-11975

    • DOI

      10.1093/nar/gkz1056

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Structural basis for oligomerization of the prokaryotic peptide transporter PepTSo22019

    • Author(s)
      Nagamura Reina、Fukuda Masahiro、Kawamoto Akihiro、Matoba Kyoko、Dohmae Naoshi、Ishitani Ryuichiro、Takagi Junichi、Nureki Osamu
    • Journal Title

      Acta Crystallographica Section F Structural Biology Communications

      Volume: 75 Issue: 5 Pages: 348-358

    • DOI

      10.1107/s2053230x19003546

    • Related Report
      2019 Annual Research Report 2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Molecular Organization and Assembly of the Export Apparatus of Flagellar Type III Secretion Systems2019

    • Author(s)
      Minamino Tohru、Kawamoto Akihiro、Kinoshita Miki、Namba Keiichi
    • Journal Title

      Current Topics in Microbiology and Immunology

      Volume: 1 Pages: 1-17

    • DOI

      10.1007/82_2019_170

    • ISBN
      9783030521226, 9783030521233
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] In Vitro Reconstitution of Functional Type III Protein Export and Insights into Flagellar Assembly2018

    • Author(s)
      Hiroyuki Terashima, Akihiro Kawamoto, Chinatsu Tatsumi, Keiichi Namba, Tohru Minamino, Katsumi Imada
    • Journal Title

      mBio

      Volume: 9(3) Issue: 3

    • DOI

      10.1128/mbio.00988-18

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] The mechanism of two-phase motility in the spirochete Leptospira : Swimming and crawling2018

    • Author(s)
      Tahara Hajime、Takabe Kyosuke、Sasaki Yuya、Kasuga Kie、Kawamoto Akihiro、Koizumi Nobuo、Nakamura Shuichi
    • Journal Title

      Science Advances

      Volume: 4 Issue: 5

    • DOI

      10.1126/sciadv.aar7975

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Leptospiral flagellar sheath protein FcpA interacts with FlaA2 and FlaB1 in Leptospira biflexa2018

    • Author(s)
      Sasaki Yuya、Kawamoto Akihiro、Tahara Hajime、Kasuga Kie、Sato Ryoichi、Ohnishi Makoto、Nakamura Shuichi、Koizumi Nobuo
    • Journal Title

      PLOS ONE

      Volume: 13 Issue: 4 Pages: e0194923-e0194923

    • DOI

      10.1371/journal.pone.0194923

    • Related Report
      2018 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] クライオ電子顕微鏡によるMycoplasma pneumoniaeの細胞接着タンパク質P1の立体構造解析2019

    • Author(s)
      川本晃大
    • Organizer
      2019年度日本マイコプラズマ学会
    • Related Report
      2019 Annual Research Report
  • [Presentation] クライオ電子顕微鏡を使った病原性細菌の運動機能や分泌機構解明への挑戦2019

    • Author(s)
      川本晃大
    • Organizer
      第92日本生化学大会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] クライオ電子顕微鏡による構造解析法チュートリアル2019

    • Author(s)
      川本晃大
    • Organizer
      CBI学会2019年大会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] クライオ電子顕微鏡を用いたタンパク質の動的機能構造解析2018

    • Author(s)
      川本晃大
    • Organizer
      大阪大学蛋白質研究所セミナー
    • Related Report
      2018 Research-status Report
  • [Presentation] Rotational symmetry structure of the bacterial flagellar motor for torque transmission revealed by cryo-EM2018

    • Author(s)
      川本晃大、宮田知子、木下実紀、南野徹、今田勝巳、加藤貴之、難波啓一
    • Organizer
      第56回日本生物物理学会
    • Related Report
      2018 Research-status Report
  • [Presentation] Structure and rotational symmetry of the rotor of the bacterial flagellar motor revealed by electron cryomicroscopy.2018

    • Author(s)
      Akihiro Kawamoto, Tomoko Miyata, Miki Kinoshita, Tohru Minamino, Katsumi Imada, Takayuki Kato, Keiichi Namba
    • Organizer
      第18回日本蛋白質科学会
    • Related Report
      2018 Research-status Report
  • [Presentation] Structure and rotational symmetry of the rotor of the bacterial flagellar motor revealed by electron cryomicroscopy.2018

    • Author(s)
      Akihiro Kawamoto, Tomoko Miyata, Miki Kinoshita, Tohru Minamino, Katsumi Imada, Takayuki Kato, Keiichi Namba
    • Organizer
      第74回日本顕微鏡学会
    • Related Report
      2018 Research-status Report
  • [Remarks] Press Release:クライオ電子顕微鏡により明らかにした細菌毒素タンパク質の膜透過機構

    • URL

      http://www.protein.osaka-u.ac.jp/%e6%9c%aa%e5%88%86%e9%a1%9e/press-release20200310_cryo

    • Related Report
      2019 Annual Research Report

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Published: 2018-04-23   Modified: 2021-02-19  

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