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Structural analysis of the interaction between spinophilin, a GPCR-signaling modulator, with GPCR

Research Project

Project/Area Number 17K07301
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Structural biochemistry
Research InstitutionGunma University

Principal Investigator

Wakamatsu Kaori  群馬大学, 大学院理工学府, 教授 (40222426)

Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywords凝集 / アミロイド / オスモライト / NMR / スピノフィリン / α2A-AR / 細胞内第三ループ / 複合体形成 / α2Aアドレナリンレセプター / HSQC / titration / 脳神経疾患 / 生体分子 / 蛋白質 / 生物物理 / 構造生物学
Outline of Final Research Achievements

By pull-down and NMR analyses, we determined the segments in spinophilin (SPL) and alpha2-adrenergic receptor (ADR) molecules that are involved in mutual interaction: 74 and 19 amino acid residues for SPL and ADR, respectively. Although ADR peptide exhibited strong tendency to form amyloid fibrils, we succeeded in preventing the amyloid formation by employing tagged ADR peptide, by adding osmolyte during concentration and titration, and by paying various precautions. [15N]SPL was titrated with non-labeled ADR and 15N-1H HSQC spectra were recorded successfully. Small but significant signal changes were observed on addition of ADR, indicating that the structural changes in the main chain of SPL is small. We are now analyzing the side chain signals of SPL by using 13C-labeled SPL, because the interaction between the two molecules is expected to be mainly of ionic nature. The strategy we developed in this study would be widely applicable for proteins that tend to form amyloid fibrils.

Academic Significance and Societal Importance of the Research Achievements

ガンや精神疾患に関与するタンパク質は他のタンパク質と相互作用することでその機能を発揮することが多い,そこでタンパク質どうしの相互作用を解析することは医学的・薬学的に重要であるが,相互作用する2種のタンパク質を混合した時に凝集したり,アミロイドを形成してしまい,相互作用を解析できないことがしばしば起こる.本研究で開発した方法はこのような問題を回避できるので,創薬の基礎的研究を推進すると期待される.

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Research-status Report
  • 2017 Research-status Report
  • Research Products

    (11 results)

All 2019 2018 2017

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

  • [Journal Article] A novel neuropilin-1 binding sequence in the human T-cell lymphotropic virus type 1 envelope glycoprotein2018

    • Author(s)
      Kusunoki H, Tanaka T, Kohno T, Matsuhashi K, Hosoda K, Wakamatsu K, Hamaguchi I
    • Journal Title

      Biochim Biophys Acta

      Volume: 1866 Issue: 4 Pages: 541-548

    • DOI

      10.1016/j.bbapap.2018.02.003

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] In-frame Val 216 -Ser 217 deletion of KIT in mild piebaldism causes aberrant secretion and SCF response2018

    • Author(s)
      Hattori Mai、Ishikawa Osamu、Oikawa Daisuke、Amano Hiroo、Yasuda Masahito、Kaira Kyoichi、Ishida-Yamamoto Akemi、Nakano Hajime、Sawamura Daisuke、Terawaki Shin-ichi、Wakamatsu Kaori、Tokunaga Fuminori、Shimizu Akira
    • Journal Title

      Journal of Dermatological Science

      Volume: 91 Issue: 1 Pages: 35-42

    • DOI

      10.1016/j.jdermsci.2018.03.012

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Journal Article] Structural basis for Ccd1 auto-inhibition in the Wnt pathway through homomerization of the DIX domain2017

    • Author(s)
      Terawaki Shin-ichi、Fujita Shohei、Katsutani Takuya、Shiomi Kensuke、Keino-Masu Kazuko、Masu Masayuki、Wakamatsu Kaori、Shibata Naoki、Higuchi Yoshiki
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 7739-7739

    • DOI

      10.1038/s41598-017-08019-5

    • NAID

      120007128850

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Identification of Candidate Genes for Generalized Tonic-Clonic Seizures in Noda Epileptic Rat2017

    • Author(s)
      Kuramoto Takashi、Voigt Birger、Nakanishi Satoshi、Kitada Kazuhiro、Nakamura Tadashi、Wakamatsu Kaori、Yoshihara Minako、Suyama Mikita、Uemura Risa、Tanaka Miyuu、Kuwamura Mitsuru、Shimizu Saki、Ohno Yukihiro、Sasa Masashi、Serikawa Tadao
    • Journal Title

      Behavior Genetics

      Volume: 47 Issue: 6 Pages: 609-619

    • DOI

      10.1007/s10519-017-9870-2

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Int'l Joint Research
  • [Presentation] アドレナリン受容体細胞内第三ループとGPCRモジュレーターSpinophilinとの相互作用解析2019

    • Author(s)
      黒川優香,葛貫絵梨奈,浦野智子,島崎安希子,河野俊之,細田和男,寺脇慎一,若松馨
    • Organizer
      第58回NMR討論会
    • Related Report
      2019 Annual Research Report
  • [Presentation] タンパク質安定化剤Non-detergent sulfobetaineによるリフォールディングの促進2018

    • Author(s)
      若松馨,小森谷美歌,浦野智子,大小原翔太,神山亮司,細田和男,寺脇慎一
    • Organizer
      第18回日本蛋白質科学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] GPCRモジュレーターspinophilinとアドレナリン受容体細胞内ループとの相互作用解析2018

    • Author(s)
      若松馨,浦野智子,島崎安希子,宮下祐美,黒川優香,細田和男,寺脇慎一
    • Organizer
      第91回日本生化学会大会
    • Related Report
      2018 Research-status Report
  • [Presentation] GABA受容体シグナリング調節因子GINIPと活性型Gαi1との相互作用解析2018

    • Author(s)
      町田結衣,小野塚樹,寺脇慎一,若松馨
    • Organizer
      日本化学会関東支部群馬地区研究交流発表会
    • Related Report
      2018 Research-status Report
  • [Presentation] Tiam1/2と活性酸素制御因子phox複合体との分子間相互作用解析2018

    • Author(s)
      田村裕貴,畑隼弥,寺脇慎一,箱嶋敏雄,若松馨
    • Organizer
      日本化学会関東支部群馬地区研究交流発表会
    • Related Report
      2018 Research-status Report
  • [Presentation] Wntシグナル伝達経路制御因子Axin-Ccd1複合体の変異体解析2018

    • Author(s)
      石渡拓也,中込蒼一朗,寺脇慎一,若松馨
    • Organizer
      日本化学会関東支部群馬地区研究交流発表会
    • Related Report
      2018 Research-status Report
  • [Presentation] GPCRシグナリング抑制因子Spinophilinとアドレナリン受容体との結合領域の絞り込みと相互作用解析2017

    • Author(s)
      浦野智子,島崎安希子,宮下祐美,細田和男,寺脇慎一,若松馨
    • Organizer
      第56回NMR討論会
    • Related Report
      2017 Research-status Report

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Published: 2017-04-28   Modified: 2021-02-19  

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