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Preparation of membrane protein-containing nanoparticles and precise observation of membrane protein dynamics

Research Project

Project/Area Number 19K22395
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionNara Institute of Science and Technology

Principal Investigator

Tsukazaki Tomoya  奈良先端科学技術大学院大学, 先端科学技術研究科, 教授 (80436716)

Project Period (FY) 2019-06-28 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2019: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Keywords膜蛋白質 / 膜タンパク質 / タンパク質
Outline of Research at the Start

細胞内で合成された蛋白質がどのように膜へと組込まれるのか,その詳細な分子メカニズムは不明であり,より詳細な解析を行うためには,1ユニット(反応最小単位)での解析が必要である。本研究では,新規ポリマーに着目した研究を進める。このポリマーは直接生体膜から膜蛋白質を生体内の脂質環境のまま取り出せ,抽出後はディスク状のナノ粒子を形成する。しかし,形成効率に問題がある。はじめに効率よく新規ポリマーを用いて膜蛋白質を抽出する系を構築する。次に蛋白質輸送に関わる必須の膜蛋白質の分子メカニズム解明に向けた活性の評価を行う。本研究による成果は,他の様々な蛋白質の詳細な解析に応用できる。

Outline of Final Research Achievements

To understand the molecular mechanism of membrane proteins in detail, a new method for monitoring a single unit of membrane protein complex was needed. In this study, we focused on membrane protein-containing nanoparticles and prepared nanodiscs containing two types of membrane proteins. In both cases, by optimizing the conditions, the membrane proteins could be observed by high-speed atomic force microscopy from two specific directions. Further studies were carried out to observe the dynamics of a single unit of membrane protein. This technique is expected to be applied to the observation of membrane protein dynamics in the future.

Academic Significance and Societal Importance of the Research Achievements

膜蛋白質の分子メカニズムを理解するには、膜に埋め込まれた状態で1分子観察する必要がある。本研究の2つの膜蛋白質の測定の成功例から、本手法はさまざまな膜タンパク質に適応可能な手法である。たとえば,次のような膜蛋白質の動態観察に有用であると考えられる。膜を隔ててシグナルを伝達する膜タンパク質では,本手法を用いることで膜の両側の構造変化を一度に確認することができ、リアルタイムでシグナル伝達の詳細を構造生物学的に理解できる。また、大きな構造変化を伴って物質を輸送するABCトランスポーターの脂質中での動態解析に適切な手法であると考えられる。

Report

(3 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • Research Products

    (17 results)

All 2021 2020 2019 Other

All Int'l Joint Research (1 results) Journal Article (8 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 7 results,  Open Access: 8 results) Presentation (6 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results) Book (1 results) Remarks (1 results)

  • [Int'l Joint Research] 味の素-ジェネチカ・リサーチ・インスティチュート社(ロシア連邦)

    • Related Report
      2020 Annual Research Report
  • [Journal Article] Proton-driven bacterial protein translocation motor SecDF2020

    • Author(s)
      塚崎 智也
    • Journal Title

      生化学

      Volume: 92 Issue: 5 Pages: 717-721

    • DOI

      10.14952/SEIKAGAKU.2020.920717

    • NAID

      40022394510

    • ISSN
      0037-1017
    • Year and Date
      2020-10-25
    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Crystal structure of a YeeE/YedE family protein engaged in thiosulfate uptake2020

    • Author(s)
      Tanaka Yoshiki、Yoshikaie Kunihito、Takeuchi Azusa、Ichikawa Muneyoshi、Mori Tomoyuki、Uchino Sayaka、Sugano Yasunori、Hakoshima Toshio、Takagi Hiroshi、Nonaka Gen、Tsukazaki Tomoya
    • Journal Title

      Science Advances

      Volume: 6 Issue: 35

    • DOI

      10.1126/sciadv.aba7637

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Reversible auto-inhibitory regulation of Escherichia coli metallopeptidase BepA for selective β-barrel protein degradation.2020

    • Author(s)
      Daimon 他7名
    • Journal Title

      Proc. Natl. Acad. Sci. USA.

      Volume: 117 Issue: 45 Pages: 27989-27996

    • DOI

      10.1073/pnas.2010301117

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structural basis of sarco/endoplasmic reticulum Ca2+-ATPase 2b regulation via transmembrane helix interplay2019

    • Author(s)
      Michio Inoue, Nanami Sakuta, Satoshi Watanabe, Yuxia Zhang, Kunihito Yoshikaie, Yoshiki Tanaka, Ryo Ushioda, Yukinari Kato, Junichi Takagi, Tomoya Tsukazaki, Kazuhiro Nagata, Kenji Inaba
    • Journal Title

      Cell Reports

      Volume: 27 Issue: 4 Pages: 1221-1230

    • DOI

      10.1016/j.celrep.2019.03.106

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Structural Basis of the Sec Translocon and YidC Revealed Through X-ray Crystallography2019

    • Author(s)
      Tsukazaki Tomoya
    • Journal Title

      The Protein Journal

      Volume: - Issue: 3 Pages: 249-261

    • DOI

      10.1007/s10930-019-09830-x

    • NAID

      120006718071

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A snapshot of membrane protein insertion2019

    • Author(s)
      Tanaka Yoshiki、Tsukazaki Tomoya
    • Journal Title

      EMBO reports

      Volume: 20 Issue: 10

    • DOI

      10.15252/embr.201949034

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] The bacterial protein YidC accelerates MPIase-dependent integration of membrane proteins2019

    • Author(s)
      Sasaki Masaru、Nishikawa Hanako、Suzuki Sonomi、Moser Michael、Huber Maria、Sawasato Katsuhiro、Matsubayashi Hideaki T.、Kumazaki Kaoru、Tsukazaki Tomoya、Kuruma Yutetsu、Nureki Osamu、Ueda Takuya、Nishiyama Ken-ichi
    • Journal Title

      Journal of Biological Chemistry

      Volume: 294 Issue: 49 Pages: 18898-18908

    • DOI

      10.1074/jbc.ra119.011248

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Real-time observation of membrane protein-embedded nanodiscs by high-speed atomic force microscopy2019

    • Author(s)
      Takamitsu Haruyama, Tomoya Tsukazaki
    • Journal Title

      野口研究所時報

      Volume: 62 Pages: 42-49

    • NAID

      40022065376

    • Related Report
      2019 Research-status Report
    • Open Access
  • [Presentation] 高速原子間顕微鏡による膜輸送装置Secの動態観察2021

    • Author(s)
      長池航,板家成良,春山隆充,塚﨑智也,内橋貴之
    • Organizer
      令和2年度中部支部講演会および 総会
    • Related Report
      2020 Annual Research Report
  • [Presentation] 高速原子間力顕微鏡によるタンパク質膜輸送装置Secの動態観察2020

    • Author(s)
      長池航,板家成良,春山隆充,塚崎智也,内橋貴之
    • Organizer
      日本生体エネルギー研究会第46回討論会
    • Related Report
      2020 Annual Research Report
  • [Presentation] Observation of Substrate Binding Sec Translocon and Structural Change of SecA with HS-AFM2020

    • Author(s)
      長池航,春山隆充,塚崎智也,内橋貴之
    • Organizer
      The 58th Annual Meeting of the BSJ
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 硫黄トランスポーターの結晶構造解析2019

    • Author(s)
      田中良樹,吉海江国仁,内野清香,塚崎智也
    • Organizer
      第16回21世紀大腸菌研究会
    • Related Report
      2019 Research-status Report
  • [Presentation] タンパク質膜透過チャネルSecトランスロコンを経由するタンパク質輸送のメカニズム2019

    • Author(s)
      塚崎智也
    • Organizer
      岩手大学放射光利用セミナー
    • Related Report
      2019 Research-status Report
    • Invited
  • [Presentation] 新奇硫黄系トランスポーターの構造と機能2019

    • Author(s)
      竹内梓, 田中良樹,吉海江 国仁, 内野清香, 高木博史, 塚﨑智也
    • Organizer
      第42回日本分子生物学会年会
    • Related Report
      2019 Research-status Report
  • [Book] 膜タンパク質工学ハンドブック2020

    • Author(s)
      浜窪隆雄, 津本浩平
    • Total Pages
      538
    • Publisher
      エヌ・ティー・エス
    • Related Report
      2020 Annual Research Report
  • [Remarks] 奈良先端大 構造生命科学 研究業績

    • URL

      https://bsw3.naist.jp/tsukazaki/publication.html

    • Related Report
      2020 Annual Research Report 2019 Research-status Report

URL: 

Published: 2019-07-04   Modified: 2022-01-27  

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