Receptor specificity and self-association of arrestin

• Project/Area Number: 16K07319
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: Kyoto University
• Principal Investigator: Imamoto Yasushi 京都大学, 理学研究科, 准教授 (80263200)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: アレスチン / G蛋白質共役型受容体 / GPCR / 自己会合 / X線溶液散乱法 / p44 / シグナル伝達 / 蛋白質 / 生体分子 / 生理学 / 脳・神経

Outline of Final Research Achievements

The interaction between rhodopsin, a typical G-protein coupled receptor, and arrestin was studied by small-angle X-ray scattering using nanodiscs. The comparison of self-association and interaction with phosphorylated rhodopsin between rod arrestin and its splice variant p44 suggested that p44 forms tetramer similar to that of arrestin. In addition, it was experimentally demonstrated that p44 and phosphorylated rhodopsin form unstable complex (pre-coupled state) in the dark, which is converted to fully coupled state by light.

Academic Significance and Societal Importance of the Research Achievements

GPCRは市販の医薬品の約半数が作用していると考えられている生理学的・薬理学的に重要な蛋白質群である。GPCRはリガンドの作用によってG蛋白質で仲介される細胞内シグナル伝達系を活性化し、リン酸化とアレスチン結合によってG蛋白質活性化能を失う。本研究で得られたアレスチンのタイプの違いによる結合様式の違いは、GPCRの活性を適切に制御して細胞が正常に機能するメカニズムの理解に寄与すると考えられる。

Research Products

• [Journal Article] Shift in Conformational Equilibrium Induces Constitutive Activity of G-Protein-Coupled Receptor, Rhodopsin. (2018)
  • Author(s): Maeda R, Hiroshima M, Yamashita T, Wada A, Sako Y, Shichida Y, Imamoto Y.
  • Journal Title: J. Phys. Chem. B Volume: 122 Issue: 18 Pages: 4838-4843
  • DOI: 10.1021/acs.jpcb.8b02819
  • NAID: 120006708207
  • Peer Reviewed / Open Access
• [Journal Article] Red-Tuning of the Channelrhodopsin Spectrum Using Long Conjugated Retinal Analogues. (2018)
  • Author(s): Shen YC, Sasaki T, Matsuyama T, Yamashita T, Shichida Y, Okitsu T, Yamano Y, Wada A, Ishizuka T, Yawo H, Imamoto Y.
  • Journal Title: Biochemistry Volume: 57 Issue: 38 Pages: 5544-5556
  • DOI: 10.1021/acs.biochem.8b00583
  • NAID: 120006708206
  • Peer Reviewed
• [Journal Article] Opn5L1 is a retinal receptor that behaves as a reverse and self-regenerating photoreceptor (2018)
  • Author(s): Sato Keita、Yamashita Takahiro、Ohuchi Hideyo、Takeuchi Atsuko、Gotoh Hitoshi、Ono Katsuhiko、Mizuno Misao、Mizutani Yasuhisa、Tomonari Sayuri、Sakai Kazumi、Imamoto Yasushi、Wada Akimori、Shichida Yoshinori
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 1255
  • DOI: 10.1038/s41467-018-03603-3
  • NAID: 120006535079
  • URL: https://tokushima-u.repo.nii.ac.jp/records/2005614
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Adaptation of cone pigments found in green rods for scotopic vision through a single amino acid mutation (2017)
  • Author(s): Kojima Keiichi、Matsutani Yuki、Yamashita Takahiro、Yanagawa Masataka、Imamoto Yasushi、Yamano Yumiko、Wada Akimori、Hisatomi Osamu、Nishikawa Kanto、Sakurai Keisuke、Shichida Yoshinori
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 114 Issue: 21 Pages: 5437-5442
  • DOI: 10.1073/pnas.1620010114
  • Peer Reviewed / Open Access
• [Journal Article] Evolutionary steps involving counterion displacement in a tunicate opsin (2017)
  • Author(s): Kojima Keiichi、Yamashita Takahiro、Imamoto Yasushi、Kusakabe Takehiro G.、Tsuda Motoyuki、Shichida Yoshinori
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 114 Issue: 23 Pages: 6028-6033
  • DOI: 10.1073/pnas.1701088114
  • Peer Reviewed / Open Access
• [Journal Article] Alternative Formation of Red-Shifted Channelrhodopsins: Noncovalent Incorporation with Retinal-Based Enamine-Type Schiff Bases and Mutated Channelopsin (2017)
  • Author(s): T. Okitsu
  • Journal Title: Chemical and Pharmaceutical Bulletin Volume: 65 Issue: 4 Pages: 356-358
  • DOI: 10.1248/cpb.c17-00054
  • NAID: 130005529782
  • ISSN: 0009-2363, 1347-5223
  • Peer Reviewed
• [Presentation] Development of Red-Shifted Channelrhodopsin Variants Using Long-Conjugated Retinal Analogues (2018)
  • Author(s): 今元 泰
  • Organizer: 第18回レチナール蛋白質国際会議
  • Int'l Joint Research / Invited
• [Presentation] Conformational Fluctuation of 115 Loop during the Photocycle of Photoactive Yellow Protein (2018)
  • Author(s): 今元 泰
  • Organizer: 日本生物物理学会第56回年会
• [Presentation] Single molecule observation of the activation dynamics of rhodopsin. (2017)
  • Author(s): Imamoto, Y.
  • Organizer: International Symposium on Biophysics of Rhodopsins
  • Int'l Joint Research / Invited
• [Presentation] Effect of dimerization on the light-induced helical rearrangement of visual rhodopsin (2017)
  • Author(s): 今元 泰、小島慧一、岡 俊彦、前田 亮、七田芳則
  • Organizer: 日本生物物理学会第55回年会
• [Presentation] Self-association of p44, a splice variant of visual rod arrestin (2016)
  • Author(s): Yasushi Imamoto, Keiichi Kojima, Toshihiko Oka, Takahiro Yamashita, Yoshinori Shichida
  • Organizer: 第54 回日本生物物理学会年会
  • Place of Presentation: つくば国際会議場
  • Year and Date: 2016-11-25
• [Presentation] Light-induced helical rearrangement of vertebrate visual rhodopsin monitored by high-angle X-ray scattering (2016)
  • Author(s): Yasushi Imamoto
  • Organizer: 17th International Conference on Retinal Proteins
  • Place of Presentation: Potsdam, Germany
  • Year and Date: 2016-10-02
  • Int'l Joint Research
• [Presentation] X線小角散乱法を用いた視細胞情報変換過程の解析 (2016)
  • Author(s): 今元 泰、小島慧一、岡 俊彦、前田 亮、七田芳則
  • Organizer: 第19回日本光生物学協会年会
  • Place of Presentation: 東京大学山上会館
  • Year and Date: 2016-07-28
• [Presentation] ロドプシン活性化の一分子ダイナミクス (2016)
  • Author(s): 今元 泰
  • Organizer: 大阪大学蛋白質研究所セミナー「膜タンパク質の構造ダイナミクス」
  • Place of Presentation: 大阪大学蛋白質研究所
  • Year and Date: 2016-05-12
  • Invited