Clarification of GPCR signaling regulation mechanism by NMR

• Project/Area Number: 20H03375
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
• Research Institution: The University of Tokyo
• Principal Investigator: Ueda Takumi 東京大学, 大学院薬学系研究科(薬学部), 准教授 (20451859)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2022: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2021: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000); Fiscal Year 2020: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
• Keywords: NMR / GPCR / 膜タンパク質 / 動的構造平衡 / Gタンパク質 / アレスチン / システム生物学 / G蛋白質 / アドレナリン受容体 / オピオイド受容体

Outline of Research at the Start

GPCRでは、異なるシグナル強度を示すリガンドの存在（薬効度）や、G蛋白質とアレスチンの一方をより強く活性化するバイアスリガンドの存在（シグナル選択性）が、創薬上重要です。本研究課題では、GPCR であるb2 アドレナリン受容体（b2AR) およびmオピオイド受容体(mOR)が、どのような構造の間を、どの程度の速度および量比で交換しているか、またそれらが薬剤の結合や活性を変化させる変異の導入に伴いどのように変化するかを明らかにして、薬効度やシグナル選択性の制御機構を解明することを目的とします。

Outline of Final Research Achievements

BMR-96122 is reportedly non-competitively bound to mu opioid receptor (mOR) and increase the signaling activity of mOR bound to the full agonist. In this study, we observed resonances from methionine residues of mOR in the presence of BMS-986122 and various ligands. Our study revealed that BMS-987122 enhances the signaling activity of mOR bound to the full agonist by perturbing the interaction between TM3 and TM6, in which signaling atctivity-related conformational dynamics was observed, and stabilizing the fully active state, in which the intraceller region adopts open conforation,

Academic Significance and Societal Importance of the Research Achievements

GPCRを標的とする医薬品は、GPCRの細胞外側に存在するポケットに結合してGPCRを活性化する。しかし、既存薬はGPCRの活性を十分に引き出すことができないことから、このポケット以外の場所に結合して作用するアロステリック薬剤の実用化が期待されている。本研究の成果により、アロステリック薬剤がGPCRに対してその構造平衡状態を変え、既存薬の限界を超えて活性を上昇させることができる可能性が示された。したがって、本研究の成果は、これまで十分な薬効を持つ医薬品が見つかっていないGPCR関連疾病に対し、十分な薬効を示す新しい医薬品開発に貢献することが期待できる。

Research Products

• [Journal Article] Amide-to-ester substitution as a stable alternative to N-methylation for increasing membrane permeability in cyclic peptides. (2023)
  • Author(s): Hosono Y, Uchida S, Shinkai M, Townsend CE, Kelly CN, Naylor MR, Lee HW, Kanamitsu K, Ishii M, Ueki R, Ueda T, Takeuchi K, Sugita M, Akiyama Y, Lokey SR, Morimoto J, Sando S.
  • Journal Title: Nat. Commun. Volume: 14 Issue: 1 Pages: 1416-1416
  • DOI: 10.1038/s41467-023-36978-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Function-Related Conformational Dynamics of GPCRs Revealed by Solution NMR (2022)
  • Author(s): UEDA Takumi、KOFUKU Yutaka、TAKEUCHI Koh、IMAI Shunsuke、SHIRAISHI Yutaro、SHIMADA Ichio
  • Journal Title: Nihon Kessho Gakkaishi Volume: 64 Issue: 4 Pages: 279-284
  • DOI: 10.5940/jcrsj.64.279
  • ISSN: 0369-4585, 1884-5576
  • Year and Date: 2022-12-15
• [Journal Article] Residue-based program of a β-peptoid twisted strand shape via a cyclopentane constraint (2022)
  • Author(s): Kim J, Kobayashi H, Yokomine M, Shiratori Y, Ueda T, Takeuchi K, Umezawa K, Kuroda D, Tsumoto K, Morimoto J, Sando S.
  • Journal Title: Org. Biomol. Chem. Volume: 20 Issue: 35 Pages: 6994-7000
  • DOI: 10.1039/d2ob01300b
  • Peer Reviewed
• [Journal Article] Activation mechanism of the μ-opioid receptor by an allosteric modulator. (2022)
  • Author(s): Kaneko S, Imai S, Asao N, Kofuku Y, Ueda T, Shimada I.
  • Journal Title: Proc. Natl. Acad. Sci. U. S. A. Volume: 119 Issue: 16
  • DOI: 10.1073/pnas.2121918119
  • Peer Reviewed / Open Access
• [Journal Article] Ultrafast water permeation through nanochannels with a densely fluorous interior surface. (2022)
  • Author(s): Itoh Y, Chen S, Hirahara R, Konda T, Aoki T, Ueda T, Shimada I, Cannon JJ, Shao C, Shiomi J, Tabata KV, Noji H, Sato K, Aida T.
  • Journal Title: Science Volume: 376 Issue: 6594 Pages: 738-743
  • DOI: 10.1126/science.abd0966
  • Peer Reviewed
• [Journal Article] Function-related dynamics of GPCRs (2022)
  • Author(s): Ueda Takumi、Imai Shunsuke、Shimada Ichio
  • Journal Title: Journal of Magnetic Resonance Volume: 336 Pages: 107164-107164
  • DOI: 10.1016/j.jmr.2022.107164
  • Peer Reviewed
• [Journal Article] Oligo(N‐methylalanine) as a Peptide‐Based Molecular Scaffold with a Minimal Structure and High Density of Functionalizable Sites (2022)
  • Author(s): Yokomine Marin、Morimoto Jumpei、Fukuda Yasuhiro、Shiratori Yota、Kuroda Daisuke、Ueda Takumi、Takeuchi Koh、Tsumoto Kouhei、Sando Shinsuke
  • Journal Title: Angewandte Chemie International Edition Volume: 61 Issue: 14 Pages: 6994-7000
  • DOI: 10.1002/anie.202200119
  • Peer Reviewed
• [Journal Article] Biphasic activation of β-arrestin 1 upon interaction with a GPCR revealed by methyl-TROSY NMR (2021)
  • Author(s): Shiraishi Yutaro、Kofuku Yutaka、Ueda Takumi、Pandey Shubhi、Dwivedi-Agnihotri Hemlata、Shukla Arun K.、Shimada Ichio
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 7158-7158
  • DOI: 10.1038/s41467-021-27482-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Function-Related Dynamics in Multi-Spanning Helical Membrane Proteins Revealed by Solution NMR (2021)
  • Author(s): Takeuchi Koh、Kofuku Yutaka、Imai Shunsuke、Ueda Takumi、Tokunaga Yuji、Toyama Yuki、Shiraishi Yutaro、Shimada Ichio
  • Journal Title: Membranes Volume: 11 Issue: 8 Pages: 604-604
  • DOI: 10.3390/membranes11080604
  • Peer Reviewed / Open Access
• [Journal Article] Structural equilibrium underlying ligand-dependent activation of b2-adrenoreceptor (2020)
  • Author(s): Shunsuke Imai, Tomoki Yokomizo, Yutaka Kofuku, Yutaro Shiraishi, Takumi Ueda, Ichio Shimada
  • Journal Title: Nature Chemical Biology Volume: 16 Issue: 4 Pages: 430-439
  • DOI: 10.1038/s41589-019-0457-5
  • Peer Reviewed
• [Journal Article] Activation of adenosine A2A receptor by lipids from docosahexaenoic acid revealed by NMR (2020)
  • Author(s): Takuya Mizumura, Keita Kondo, Masatoshi Kurita, Yutaka Kofuku, Mei Natsume, Shunsuke Imai, Yutaro Shiraishi, Takumi Ueda, Ichio Shimada
  • Journal Title: Science Advances Volume: 6 Issue: 12
  • DOI: 10.1126/sciadv.aay8544
  • Peer Reviewed / Open Access
• [Presentation] アデノシンA2A受容体とリガンドの滞在時間を規定する構造基盤の解明 (2022)
  • Author(s): 上田 卓見
  • Organizer: 第61回NMR討論会
• [Presentation] Clarification of the GPCR signaling mechanism based on the function-related conformational dynamics (2022)
  • Author(s): 上田 卓見
  • Organizer: 第61回NMR討論会
  • Invited
• [Remarks]
  • URL: https://biophys.f.u-tokyo.ac.jp/
• [Remarks]
  • URL: https://biophys.f.u-tokyo.ac.jp/
• [Remarks]
  • URL: http://ishimada.f.u-tokyo.ac.jp/public_html/index_j.html