Structural basis of biased signaling mediated by pharmacologically important GPCRs

• Project/Area Number: 15K18843
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Physical pharmacy
• Research Institution: The University of Tokyo
• Principal Investigator: KOFUKU Yutaka 東京大学, 大学院薬学系研究科(薬学部), 特任助教 (80737940)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 構造生物学 / NMR / 膜タンパク質 / シグナル伝達 / GPCR

Outline of Final Research Achievements

G-protein-coupled receptor (GPCR) ligands impart differing degrees of signaling in the G-protein and arrestin pathways, in phenomena called "biased signaling". However, the mechanism underlying the biased signaling of GPCRs is still unclear, although crystal structures of GPCRs bound to the G protein or arrestin are available. In this study, we observed the NMR signals from methionine residues of pharmacologically important GPCRs in the balanced- and biased-ligand-bound states. We found that the intracellular cavity of GPCRs exist in an equilibrium between closed and multiple open conformations, and that the population of each open conformation determines the G-protein- and arrestin-mediated signaling levels in each ligand-bound state.

Research Products

• [Journal Article] Conductance of P2X4 receptor is determined by conformational equilibrium in the transmembrane region (2016)
  • Author(s): Minato Y, Suzuki S , Hara T, Kofuku Y, Kasuya G, Fujiwara Y, Igarashi S, Suzuki E, Nureki O, Hattori M, Ueda T, Shimada I
  • Journal Title: Proc. Natl. Acad. Sci. Volume: 発行中 Issue: 17 Pages: 4741-4746
  • DOI: 10.1073/pnas.1600519113
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Utilization of paramagnetic relaxation enhancements for structural analysis of actin-binding proteins in complex with actin. (2016)
  • Author(s): Huang S, Umemoto R, Tamura Y, Kofuku Y, Uyeda TQP, Nishida N, Shimada I
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 33690-33690
  • DOI: 10.1038/srep33690
  • Peer Reviewed / Open Access
• [Journal Article] Identification of a Conformational Equilibrium That Determines the Efficacy and Functional Selectivity of the μ-Opioid Receptor. (2015)
  • Author(s): Junya Okude, Takumi Ueda, Yutaka Kofuku, Motohiko Sato, Naoyuki Nobuyama, Keita Kondo, Yutaro Shiraishi, Takuya Mizumura, Kento Onishi, Mei Natsume, Masahiro Maeda, Hideki Tsujishita, Takefumi Kuranaga, Masayuki Inoue, Ichio Shimada
  • Journal Title: Angewandte Chemie International Edition Volume: 21 Issue: 52 Pages: 15771-15776
  • DOI: 10.1002/anie.201508794
  • Peer Reviewed / Open Access
• [Journal Article] Elucidation of the CCR1- and CCR5-binding modes of MIP-1α by application of an NMR spectra reconstruction method to the transferred cross-saturation experiments. (2015)
  • Author(s): Chie Yoshiura, Takumi Ueda, Yutaka Kofuku, Masahiko Matsumoto, Junya Okude, Keita Kondo, Yutaro Shiraishi, Ichio Shimada
  • Journal Title: Journal of Biomolecular NMR Volume: 63 Issue: 4 Pages: 333-340
  • DOI: 10.1007/s10858-015-9992-x
  • Peer Reviewed / Open Access
• [Journal Article] Development of a method for reconstruction of crowded NMR spectra from undersampled time-domain data (2015)
  • Author(s): Ueda T, Yoshiura C, Matsumoto M, Kofuku Y, Okude J, Kondo K, Shiraishi Y, Takeuchi K, Shimada I
  • Journal Title: J. Biomol. NMR Volume: 印刷中 Issue: 1 Pages: 31-41
  • DOI: 10.1007/s10858-015-9908-9
  • Peer Reviewed / Open Access
• [Presentation] 細胞遊走因子、ケモカイン受容体の動的構造研究と創薬研究への展望 (2017)
  • Author(s): 幸福 裕
  • Organizer: 第87回日本衛生学会学術総会
  • Place of Presentation: シーガイアコンベンションセンター（宮崎県宮崎市）
  • Year and Date: 2017-03-26
  • Invited
• [Presentation] Conductance of P2X4 purinergic receptor is determined by conformational equilibrium in the transmembrane region (2017)
  • Author(s): Takumi Ueda, Tomoaki Hara, Yuichi Minato, Shiho Suzuki, Yutaka Kofuku, Go Kasuya, Yuichiro Fujiwara, Shunsuke Igarashi, Ei-ichiro Suzuki, Osamu Nureki, Motoyuki Hattori, Ichio Shimada
  • Organizer: 58th Experimental Nuclear Magnetic Resonance Conference
  • Place of Presentation: Pacific Grove（アメリカ合衆国）
  • Year and Date: 2017-03-26
  • Int'l Joint Research / Invited
• [Presentation] Identification of a Conformational Equilibrium That Determines the Efficacy and Functional Selectivity of the μ-Opioid Receptor (2017)
  • Author(s): Yutaro Shiraishi, Junya Okude, Takumi Ueda, Yutaka Kofuku, Motohiko Sato, Naoyuki Nobuyama, Keita Kondo, Takuya Mizumura, Mei Natsume, Masahiro Maeda, Hideki Tsujishita, Takefumi Kuranaga, Masayuki Inoue, Ichio Shimada
  • Organizer: 58th Experimental Nuclear Magnetic Resonance Conference
  • Place of Presentation: Pacific Grove（アメリカ合衆国）
  • Year and Date: 2017-03-26
  • Int'l Joint Research
• [Presentation] Functional dynamics of deuterated β2-adrenergic receptor in lipid bilayers revealed by NMR spectroscopy (2017)
  • Author(s): Mei Natsume, Yutaka Kofuku, Takumi Ueda, Junya Okude, Yutaro Shiraishi, Keita Kondo, Takuya Mizumura, Shiho Suzuki, Ichio Shimada
  • Organizer: 58th Experimental Nuclear Magnetic Resonance Conference
  • Place of Presentation: Pacific Grove（アメリカ合衆国）
  • Year and Date: 2017-03-26
  • Int'l Joint Research
• [Presentation] 昆虫細胞発現系における高度重水素標識を用いた脂質二重膜中のGタンパク質共役型受容体の動的構造解析 (2016)
  • Author(s): 幸福 裕、上田 卓見、奥出 順也、白石 勇太郎、近藤 啓太、水村 拓也、鈴木 志歩、嶋田 一夫
  • Organizer: 第55回NMR討論会
  • Place of Presentation: 広島国際会議場（広島県広島市）
  • Year and Date: 2016-11-16
• [Presentation] Identification of a Conformational Equilibrium That Determines the Efficacy and Functional Selectivity of the μ-Opioid Receptor (2016)
  • Author(s): Junya Okude, Takumi Ueda, Yutaka Kofuku, Motohiko Sato, Naoyuki Nobuyama, Keita Kondo, Yutaro Shiraishi, Takuya Mizumura, Mei Natsume, Masahiro Maeda, Hideki Tsujishita, Takefumi Kuranaga, Masayuki Inoue, Ichio Shimada
  • Organizer: XXVIIth International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-08-21
  • Int'l Joint Research
• [Presentation] Development of an NMR reconstruction method to elucidate the CCR1- and CCR5-bincing modes of MIP-1a (2016)
  • Author(s): Takumi Ueda, Chie Yoshiura, Yutaka Kofuku, Masahiko Matsumoto, Junya Okude, Keita Kondo, Yutaro Shiraishi, Koh Takeuchi, Ichio Shimada
  • Organizer: XXVIIth International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-08-21
  • Int'l Joint Research
• [Presentation] Conductance of P2X4 purinergic receptor is determined by conformational equilibrium in the transmembrane region (2016)
  • Author(s): Tomoaki Hara, Yuichi Minato, Shiho Suzuki, Yutaka Kofuku, Go Kasuya, Yuichiro Fujiwara, Shunsuke Igarashi, Ei-ichiro Suzuki, Osamu Nureki, Motoyuki Hattori, Takumi Ueda, Ichio Shimada
  • Organizer: XXVIIth International Conference on Magnetic Resonance in Biological Systems
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-08-21
  • Int'l Joint Research
• [Presentation] δオピオイド受容体（δOR）の活性制御機構の解明 (2016)
  • Author(s): 佐藤 元彦、奥出 順也、延山 尚幸、上田 卓見、幸福 裕、倉永 健史、井上 将行、嶋田 一夫
  • Organizer: 日本薬学会第136年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-03-26
• [Presentation] SDF-1との二段階結合によるCXCR4のシグナル伝達機構の解明 (2016)
  • Author(s): 大西 健人、幸福 裕、塚本 宇信、鈴木 志歩、上田 卓見、嶋田 一夫
  • Organizer: 日本薬学会第136年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-03-26
• [Presentation] NMR法を用いた脂質二重膜中のGPCRの動的構造解析 (2015)
  • Author(s): 幸福 裕、上田 卓見、奥出 順也、白石 勇太郎、近藤 啓太、鈴木 志歩、嶋田 一夫
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学 角間キャンパス 自然科学本館（石川県金沢市）
  • Year and Date: 2015-09-13
  • Invited
• [Presentation] リガンドとの相互作用に伴う膜蛋白質の動的立体構造変化 (2015)
  • Author(s): 上田 卓見、幸福 裕、嶋田 一夫
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学 角間キャンパス 自然科学本館（石川県金沢市）
  • Year and Date: 2015-09-13
  • Invited
• [Presentation] 区分同位体標識を用いたβ2-アドレナリン受容体のリン酸化のNMR解析 (2015)
  • Author(s): 白石 勇太郎、幸福 裕、上田 卓見、夏目 芽衣、岩井 秀夫、嶋田 一夫
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学 角間キャンパス 自然科学本館（石川県金沢市）
  • Year and Date: 2015-09-13
• [Presentation] 脂質によるGPCRの活性制御機構の解明 (2015)
  • Author(s): 水村 拓也、近藤 啓太、上田 卓見、幸福 裕、嶋田 一夫
  • Organizer: 第53回日本生物物理学会年会
  • Place of Presentation: 金沢大学 角間キャンパス 自然科学本館（石川県金沢市）
  • Year and Date: 2015-09-13
• [Remarks] 東京大学大学院薬学系研究科生命物理化学教室ホームページ
  • URL: http://ishimada.f.u-tokyo.ac.jp/public_html/index_j.html