Structure determination of GPCRs using CryoEM.

• Project/Area Number: 17K19349
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Research Field: Molecular and Genome biology and related fields
• Research Institution: Kyoto University
• Principal Investigator: Shimamura Tatsuro 京都大学, 医学研究科, 特定講師 (90391979)
• Project Period (FY): 2017-06-30 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000); Fiscal Year 2018: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2017: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: 構造解析 / GPCR / 電子顕微鏡 / G蛋白質共役型受容体

Outline of Final Research Achievements

G protein-coupled receptors (GPCRs) are important drug targets. By binding a Fab fragment, we tried to determine the structures of GPCRs by electron microscopy. Unfortunately, the antibody was observed to be dissociated from GPCRs during the cryo-electron microscopy grid preparation. As a result, we have not succeeded to determine the structure of GPCRs by electron microscopy.

Academic Significance and Societal Importance of the Research Achievements

多様な生命現象に関わっているGPCRは、機能的にも創薬ターゲットとしても重要な膜タンパク質である。従来のX線結晶構造解析の手法に加え、クライオ電子顕微鏡法でもGPCRの構造解析が可能になると、GPCRの立体構造が解明できる可能性が高まり、GPCRの機能の解析や、合理的な創薬に役立つ。本研究計画では、実際にクライオ電子顕微鏡法でGPCRの構造を決定するには至らなかったが、研究の過程で培った経験は、電子顕微鏡を用いた膜タンパク質の構造解析に役立つと考えられる。

Research Products

• [Journal Article] The Crystal Structure of Angiotensin II Type 2 Receptor with Endogenous Peptide Hormone (2020)
  • Author(s): Asada Hidetsugu、Inoue Asuka、Ngako Kadji Francois Marie、Hirata Kunio、Shiimura Yuki、Im Dohyun、Shimamura Tatsuro、Nomura Norimichi、Iwanari Hiroko、Hamakubo Takao、Kusano-Arai Osamu、Hisano Hiromi、Uemura Tomoko、Suno Chiyo、Aoki Junken、Iwata So
  • Journal Title: Structure Volume: 28 Issue: 4 Pages: 418-425.e4
  • DOI: 10.1016/j.str.2019.12.003
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] High-viscosity sample-injection device for serial femtosecond crystallography at atmospheric pressure (2019)
  • Author(s): Shimazu Yoshiaki、Tono Kensuke、Tanaka Tomoyuki、Yamanaka Yasuaki、Nakane Takanori、Mori Chihiro、Terakado Kimura Kanako、Fujiwara Takaaki、Sugahara Michihiro、Tanaka Rie、Doak R. Bruce、Shimamura Tatsuro、Iwata So、Nango Eriko、Yabashi Makina
  • Journal Title: Journal of Applied Crystallography Volume: 52 Issue: 6 Pages: 1280-1288
  • DOI: 10.1107/s1600576719012846
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structural basis for the substrate recognition of aminoglycoside 7′′-phosphotransferase-Ia from Streptomyces hygroscopicus (2019)
  • Author(s): Takenoya Mihoko、Shimamura Tatsuro、Yamanaka Ryuji、Adachi Yuya、Ito Shinsaku、Sasaki Yasuyuki、Nakamura Akira、Yajima Shunsuke
  • Journal Title: Acta Crystallographica Section F Structural Biology Communications Volume: 75 Issue: 9 Pages: 599-607
  • DOI: 10.1107/s2053230x19011105
  • NAID: 120007133009
  • Peer Reviewed / Open Access
• [Journal Article] 5-HT2A受容体の構造から解明された薬剤の受容体選択機構 (2019)
  • Author(s): 島村達郎
  • Journal Title: 医学のあゆみ Volume: 270 Pages: 1152-1153
• [Journal Article] Structures of the 5-HT2A receptor in complex with the antipsychotics risperidone and zotepine (2019)
  • Author(s): Kimura Kanako Terakado、Asada Hidetsugu、Inoue Asuka、Kadji Francois Marie Ngako、Im Dohyun、Mori Chihiro、Arakawa Takatoshi、Hirata Kunio、Nomura Yayoi、Nomura Norimichi、Aoki Junken、Iwata So、Shimamura Tatsuro
  • Journal Title: Nature Structural & Molecular Biology Volume: 26 Issue: 2 Pages: 121-128
  • DOI: 10.1038/s41594-018-0180-z
  • Peer Reviewed
• [Journal Article] Crystal structure of the human angiotensin II type 2 receptor bound to an angiotensin II analog (2018)
  • Author(s): Asada H, Horita S, Hirata K, Shiroishi M, Shiimura Y, Iwanari H, Hamakubo T, Shimamura T, Nomura N, Kusano-Arai O, Uemura T, Suno C, Kobayashi T, Iwata S.
  • Journal Title: Nature Structural and Molecular Biology Volume: 25 Issue: 7 Pages: 570-576
  • DOI: 10.1038/s41594-018-0079-8
  • Peer Reviewed
• [Journal Article] Structure, Substrate Recognition, and Mechanism of the Na+-Hydantoin Membrane Transport Protein, Mhp1 (2018)
  • Author(s): Jackson Scott M.、Ivanova Ekaterina、Calabrese Antonio N.、Polyakova Anna、Sharples David J.、Shimamura Tatsuro、Cameron Alexander D.、Henderson Peter J. F., et al.
  • Journal Title: Encyclopedia of Biophysics Volume: 1 Pages: 1-12
  • DOI: 10.1007/978-3-642-35943-9_10091-1
  • ISBN: 9783642359439
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Structure of orexin receptor determined using X-ray free electron laser. (2018)
  • Author(s): Shimamura T.
  • Organizer: AsCA 2018/Crystal32
  • Int'l Joint Research / Invited
• [Presentation] Structural insights into antipsychotics binding modes in dopamine D2-class receptors (2018)
  • Author(s): Dohyun Im, Tatsuro Shimamura, et al.
  • Organizer: Keystone Symposia: GPCR Structure and Function
  • Int'l Joint Research
• [Presentation] A first novel GPCR structure determined at SACLA (2017)
  • Author(s): Dohyun Im, Tatsuro Shimamura, So Iwata
  • Organizer: International Symposium on Frontiers in structural biology for Drug Development
  • Int'l Joint Research