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Activation mechanism of G protein-coupled receptors by X-ray free electron lasers

Research Project

Project/Area Number 18H02394
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43020:Structural biochemistry-related
Research InstitutionTohoku University (2020)
Institute of Physical and Chemical Research (2018-2019)

Principal Investigator

Nango Eriko  東北大学, 多元物質科学研究所, 教授 (90376947)

Co-Investigator(Kenkyū-buntansha) 浅田 秀基  京都大学, 医学研究科, 特定講師 (20399041)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
Fiscal Year 2020: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2019: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2018: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000)
KeywordsX線結晶構造解析 / Gタンパク質共役型受容体 / X線自由電子レーザー / シリアルフェムト秒結晶構造解析 / ロドプシン / X線自由電子レーザー
Outline of Final Research Achievements

We conducted this research to analyze dynamic structures of visual rhodopsin, which is one of G protein-coupled receptors, using X-ray free electron lasers (XFEL). So far, time-resolved crystallography using XFEL has enabled to visualize dynamic structures of several light-sensitive proteins. However, it has been challenging to measure visual rhodopsin crystals because the crystal is damaged by very weak light. It has been also challenging to obtain rhodopsin crystals diffracting with high spatial resolution. We developed a new experimental device for time-resolved serial femtosecond crystallography to transport samples discontinuously for less light contamination. We performed pump-probe type time-resolved experiments with photo-sensitive samples using the device and observed structural changes with time. In addition, various expression systems of visual rhodopsin have been examined, and crystals were obtained under several conditions.

Academic Significance and Societal Importance of the Research Achievements

従来、活性化状態に至る途中構造など動的構造を原子分解能で得ることは困難であった。新たな放射光源であるX線自由電子レーザー(XFEL)を用いると動的構造を可視化することが可能であるが、視覚ロドプシンのような取り扱いが難しい試料の測定に適した装置や技術は達成されていなかった。本課題ではXFEL実験用装置開発に取り組み、新たな測定方法を確立した。こうした技術は他試料の動的構造決定においても貢献すると期待される。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Annual Research Report
  • 2018 Annual Research Report
  • Research Products

    (12 results)

All 2021 2020 2019 2018

All Journal Article (4 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 4 results,  Open Access: 4 results) Presentation (8 results) (of which Int'l Joint Research: 4 results,  Invited: 7 results)

  • [Journal Article] Structure of the dopamine D2 receptor in complex with the antipsychotic drug spiperone2020

    • Author(s)
      Im Dohyun、Inoue Asuka、Fujiwara Takaaki、Nakane Takanori、Yamanaka Yasuaki、Uemura Tomoko、Mori Chihiro、Shiimura Yuki、Kimura Kanako Terakado、Asada Hidetsugu、Nomura Norimichi、Tanaka Tomoyuki、Yamashita Ayumi、Nango Eriko、Tono Kensuke、Kadji Francois Marie Ngako、Aoki Junken、Iwata So、Shimamura Tatsuro
    • Journal Title

      Nature Communications

      Volume: 11 Issue: 1 Pages: 1-11

    • DOI

      10.1038/s41467-020-20221-0

    • NAID

      120006943309

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] A tool for visualizing protein motions in time-resolved crystallography2020

    • Author(s)
      Wickstrand Cecilia、Katona Gergely、Nakane Takanori、Nogly Przemyslaw、Standfuss Joerg、Nango Eriko、Neutze Richard
    • Journal Title

      Structural Dynamics

      Volume: 7 Issue: 2 Pages: 024701-024701

    • DOI

      10.1063/1.5126921

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] High-viscosity sample-injection device for serial femtosecond crystallography at atmospheric pressure2019

    • Author(s)
      Shimazu Yoshiaki、Tono Kensuke、Tanaka Tomoyuki、Yamanaka Yasuaki、Nakane Takanori、Mori Chihiro、Terakado Kimura Kanako、Fujiwara Takaaki、Sugahara Michihiro、Tanaka Rie、Doak R. Bruce、Shimamura Tatsuro、Iwata So、Nango Eriko、Yabashi Makina
    • Journal Title

      Journal of Applied Crystallography

      Volume: 52 Issue: 6 Pages: 1280-1288

    • DOI

      10.1107/s1600576719012846

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Pump-Probe Time-Resolved Serial Femtosecond Crystallography at SACLA: Current Status and Data Collection Strategies2019

    • Author(s)
      Nango Eriko、Kubo Minoru、Tono Kensuke、Iwata So
    • Journal Title

      Applied Sciences

      Volume: 9 Issue: 24 Pages: 5505-5505

    • DOI

      10.3390/app9245505

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Presentation] Time-resolved serial femtosecond crystallography of light-sensitive proteins2021

    • Author(s)
      Eriko Nango
    • Organizer
      Belgian Crystallography Symposium BCS-11
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Measurement Systems for Biomolecular Movies using X-ray Free Electron Lasers2021

    • Author(s)
      Eriko Nango
    • Organizer
      SACLA Users' Meeting 2021
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Protein structural dynamics revealed by molecular movie analysis2021

    • Author(s)
      Eriko Nango
    • Organizer
      RIKEN BDR Symposium 2021 “Structuring Biosystems: FunctionsEmerging from Molecules”
    • Related Report
      2020 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 高速分子動画:タンパク質が機能する際の構造変化を可視化する2020

    • Author(s)
      南後 恵理子
    • Organizer
      第43回日本分子生物学会年会シンポジウム 拡大する構造生物学 Expanding Structural Biology
    • Related Report
      2020 Annual Research Report
    • Invited
  • [Presentation] A molecular movie of structural changes in the light-driven proton pump bacteriorhodopsin2019

    • Author(s)
      Eriko Nango
    • Organizer
      2019 ESP-IUPB World Congress
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] X線自由電子レーザーによるロドプシン類の分子動画解析2019

    • Author(s)
      南後恵理子
    • Organizer
      ISSP ワークショップ「レチナールタンパク質の光機能発現の物理と化学」
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] X 線自由電子レーザーによるタンパク質分子動画撮影2019

    • Author(s)
      南後恵理子
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] XFEL分子動画が捉えたタンパク質構造変化2018

    • Author(s)
      南後恵理子
    • Organizer
      日本蛋白質科学会年会
    • Related Report
      2018 Annual Research Report
    • Invited

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Published: 2018-04-23   Modified: 2022-01-27  

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