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Dynamical structure analysis of intrinsically disordered proteins by neutron scattering using segment deuteration method

Research Project

Project/Area Number 18H02391
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 InstitutionYokohama City University

Principal Investigator

SATO Mamoru  横浜市立大学, 生命医科学研究科, 教授 (60170784)

Co-Investigator(Kenkyū-buntansha) 小田 隆  立教大学, 理学部, 助教 (00573164)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,680,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥4,080,000)
Fiscal Year 2020: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2019: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥10,270,000 (Direct Cost: ¥7,900,000、Indirect Cost: ¥2,370,000)
Keywords天然変性タンパク質 / 中性子小角散乱 / セグメント重水素化
Outline of Final Research Achievements

In the dynamical structural analysis of proteins by neutron scattering (SANS), neutron facilities overseas were not available at all due to the COVID-19 pandemic. So, X-ray scattering (SAXS) data collected prior to SANS experiment were used for preliminary analysis. The analysis was performed by ensemble optimization method. As a result, the structure of the full-length Hef protein in solution could be well reproduced as an ensemble of six structures with different conformations.
Furthermore, we have established a method for efficient preparation of polypeptide chain ligation by using split intein, NpuDnaE. In addition, a sample preparation protocol in which the deuteration ratio was controlled and a protocol for evaluating the deuteration ratio were also established.

Academic Significance and Societal Importance of the Research Achievements

タンパク質の動的構造の情報は創薬の分野で創薬候補化合物のヒット率の向上に大きく寄与するものと期待され、わが国の国家プロジェクトとして、京/富嶽コンピューターを利用した分子動力学計算で予測されてきた。しかし、これはあくまでもin silicoのシミュレーションの世界の話で、実験データに基づいたタンパク質の動的構造解析法の開発が長く待たれていた。その意味で、当該研究は学術的な意義とともに社会的意義も非常に大きいと期待される。

Report

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

    (19 results)

All 2020 2019 2018

All Journal Article (8 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 6 results,  Open Access: 5 results) Presentation (10 results) (of which Int'l Joint Research: 1 results,  Invited: 5 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Serine 298 Phosphorylation in Linker 2 of UHRF1 Regulates Ligand-Binding Property of Its Tandem Tudor Domain2020

    • Author(s)
      Kori Satomi、Jimenji Tomohiro、Ekimoto Toru、Sato Miwa、Kusano Fumie、Oda Takashi、Unoki Motoko、Ikeguchi Mitsunori、Arita Kyohei
    • Journal Title

      Journal of Molecular Biology

      Volume: 432 Issue: 14 Pages: 4061-4075

    • DOI

      10.1016/j.jmb.2020.05.006

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Structural and dynamics analysis of intrinsically disordered proteins by high-speed atomic force microscopy2020

    • Author(s)
      Kodera Noriyuki、Noshiro Daisuke、Dora Sujit K.、Mori Tetsuya、Habchi Johnny、Blocquel David、Gruet Antoine、Dosnon Marion、Salladini Edoardo、Bignon Christophe、Fujioka Yuko、Oda Takashi、Noda Nobuo N.、Sato Mamoru、Lotti Marina、Mizuguchi Mineyuki、Longhi Sonia、Ando Toshio
    • Journal Title

      Nature Nanotechnology

      Volume: 16 Issue: 2 Pages: 181-189

    • DOI

      10.1038/s41565-020-00798-9

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Dynamics of proteins with different molecular structures under solution condition2020

    • Author(s)
      Inoue Rintaro、Oda Takashi、Nakagawa Hiroshi、Tominaga Taiki、Saio Tomohide、Kawakita Yukinobu、Shimizu Masahiro、Okuda Aya、Morishima Ken、Sato Nobuhiro、Urade Reiko、Sato Mamoru、Sugiyama Masaaki
    • Journal Title

      Scientific Reports

      Volume: 10 Issue: 1 Pages: 21678-21678

    • DOI

      10.1038/s41598-020-78311-4

    • NAID

      120007193758

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structure of HIRAN Domain of Human HLTF Bound to Duplex DNA Provides Structural Basis for DNA Unwinding to Initiate Replication Fork Regression2020

    • Author(s)
      Asami Hishiki, Mamoru Sato, Hiroshi Hashimoto
    • Journal Title

      J. Biochem

      Volume: - Issue: 6 Pages: 597-602

    • DOI

      10.1093/jb/mvaa008

    • NAID

      40022260390

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Structure of the UHRF1 Tandem Tudor Domain Bound to a Methylated Non-histone Protein, LIG1, Reveals Rules for Binding and Regulation2019

    • Author(s)
      Kori, S. Ferry, L. Matano, S. Jimenji, T. Kodera, N. Tsusaka, T. Matsumura, R. Oda, T. Sato, M. Dohmae, N. Ando, T. Shinkai, Y. Defossez, P. A. Arita, K.
    • Journal Title

      Structure

      Volume: 27 Issue: 3 Pages: 1-12

    • DOI

      10.1016/j.str.2018.11.012

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Cooperative DNA Binding of the Plasmid Partitioning Protein TubR from the Bacillus cereus pXO1 Plasmid2018

    • Author(s)
      I. Hayashi, T. Oda, M. Sato, S. Fuchigami
    • Journal Title

      J. Mol. Biol.

      Volume: 430 Issue: 24 Pages: 5015-5028

    • DOI

      10.1016/j.jmb.2018.11.001

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] 細胞外での分子修飾(細胞外基質/その他)シトルリン化2018

    • Author(s)
      佐藤 衛
    • Journal Title

      生体の科学(増大特集:タンパク質・核酸の分子修飾)

      Volume: 69 Pages: 520-521

    • Related Report
      2018 Annual Research Report
  • [Journal Article] 天然変性蛋白質-蛋白質の構造・機能研究の新しいターゲット2018

    • Author(s)
      佐藤 衛
    • Journal Title

      医学のあゆみ(第5土曜特集:蛋白質代謝医学-構造・機能の研究から臨床応用まで)

      Volume: 267 Pages: 954-958

    • Related Report
      2018 Annual Research Report
  • [Presentation] Why we need deuteration in the field of life science now2019

    • Author(s)
      佐藤 衛
    • Organizer
      第4回 重水素材料研究会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] 天然変性タンパク質の構造研究とLLPSの構造基盤構築への展望2019

    • Author(s)
      佐藤 衛
    • Organizer
      第4回LLPS研究会・ASUKA若手交流会2019
    • Related Report
      2019 Annual Research Report
    • Invited
  • [Presentation] The dynamic structure and function of intrinsically disordered region of Hef that is associated with a DNA repair2019

    • Author(s)
      T. Oda, A. Sekino, A. Murakami1, R. Oi, M. Yoneyama, N. Kodera, T. Ando, T. Konuma, K. Sugase, T. Oroguchi, R. Inoue, M. Sugiyama, S. Ishino, Y. Ishino, and M. Sato
    • Organizer
      The 3rd Asia-Oceania Conference on Neutron Scattering 2019 (AOCNS 2019)
    • Related Report
      2019 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] DNA修復にかかわるHefの天然変性領域の揺らいだ構造と機能2019

    • Author(s)
      小田 隆、関野絢子、村上綾香、大井里香、米山真紀、古寺哲幸、安藤敏夫、小沼 剛、菅瀬謙治、苙口友隆、井上倫太郎、杉山正明、石野園子、石野良純、佐藤 衛
    • Organizer
      第4回LLPS研究会・ASUKA若手交流会2019
    • Related Report
      2019 Annual Research Report
  • [Presentation] The structure and function of intrinsically disordered region of Hef that is associated with a DNA repair(DNA修復にかかわるHefの天然変性領域の構造と機能)2019

    • Author(s)
      T. Oda, A. Sekino, A. Murakami, R. Oi, M. Yoneyama, N. Kodera, T. Ando, T. Konuma, K. Sugase, T. Oroguchi, S. Ishino, Y. Ishino, M. Sato
    • Organizer
      第57回日本生物物理学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] SEX-SAXSによるLDLRファミリーのpH依存的な構造変化の検証2019

    • Author(s)
      塩澤亜紀、飯田麻生、小田 隆、佐藤 衛、禾 晃和
    • Organizer
      第19回日本蛋白質科学会年会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Protein deuteration for small angle neutron scattering2019

    • Author(s)
      小田 隆、関野絢子、米山真紀、村上綾香、苙口友隆、浴本 亨、池口満徳、杉山正明、石野良純、佐藤 衛
    • Organizer
      J-PARC Workshop2018(Deuterium Labeling Study for Neutron Science)
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 中性子溶液散乱を基軸とした蛋白質の構造・ダイナミクス解析への挑戦2018

    • Author(s)
      杉山正明、中川 洋、斉尾智英、矢木真穂、苙口友隆、小田 隆、井上倫太郎、佐藤信浩、守島 健、日野正裕、川北至信、高田愼一、富永大輝、佐藤 衛
    • Organizer
      日本中性子科学会第18回年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] SANSと計算科学による天然変性タンパク質の動的構造と機能の解明2018

    • Author(s)
      小田 隆、関野絢子、米山真紀、浴本 亨、池口満徳、苙口友隆、杉山正明、石野良純、佐藤 衛
    • Organizer
      日本中性子科学会第18回年会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 溶液散乱と計算科学で明らかにする天然変性タンパク質の構造と機能2018

    • Author(s)
      小田 隆、関野絢子、米山真紀、村上綾香、苙口友隆、浴本 亨、池口満徳、杉山正明、石野良純、佐藤 衛
    • Organizer
      PF研究会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Patent(Industrial Property Rights)] 新規抗PAD4抗体2018

    • Inventor(s)
      佐藤 衛、山田 道之、金澤 智
    • Industrial Property Rights Holder
      佐藤 衛、山田 道之、金澤 智
    • Industrial Property Rights Type
      特許
    • Filing Date
      2018
    • Acquisition Date
      2020
    • Related Report
      2019 Annual Research Report

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

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