立体構造に基づくGPCRとリガンドの相互作用の解明

• Project Area: Chemical Biology using bioactive natural products as specific ligands: identification of molecular targets and regulation of bioactivity
• Project/Area Number: 26102725
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Kyoto University
• Principal Investigator: 島村 達郎 京都大学, 医学(系)研究科(研究院), 講師 (90391979)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2015: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2014: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: GPCR / リガンド

Outline of Annual Research Achievements

GPCRの活性型構造は、agonistを結合させただけでは不安定でることから構造解析は難しく、agonistの結合様式に関する構造情報は限られている。本年度は、昨年度までに作製したagonistの親和性を高める構造認識抗体を結合させ、agonistが結合した状態の受容体の構造解析に成功した。構造解析の結果、結合させたagonistの電子密度は良好で、このagonistの結合様式の詳細が解明できた。また、類似の手法を他の膜蛋白質の構造解析に応用することにも成功した。また、最近になり、X線結晶構造解析に次世代X線源であるX線自由電子レーザーが利用できるようになった。我々は、X線自由電子レーザーを受容体の結晶構造解析に利用するための測定系を立ち上げ、いくつかの受容体の構造解析に成功した。そのうちの一種では、従来の放射光を用いて測定したデータより、リガンド周囲の電子密度が改善され、より詳細なリガンドの結合様式が解明できた。本研究で得られた成果は、他の受容体や膜蛋白質とリガンドの結合様式の詳細を解明するための膜蛋白質構造解析技術基盤の確立にも役立った。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Haem-dependent dimerization of PGRMC1/Sigma-2 receptor facilitates cancer proliferation and chemoresistance. (2016)
  • Author(s): Kabe Y, Nakane T, Koike I, Yamamoto T, Sugiura Y, Harada E, Sugase K, Shimamura T, Ohmura M, Muraoka K, Yamamoto A, Uchida T, Iwata S, Yamaguchi Y, Krayukhina E, Noda M, Handa H, Ishimori K, Uchiyama S, Kobayashi T, Suematsu M.
  • Journal Title: Nature Communications Volume: 7 Issue: 1 Pages: 11030-11030
  • DOI: 10.1038/ncomms11030
  • NAID: 130007529314
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Native sulfur/chlorine SAD phasing for serial femtosecond crystallography. (2015)
  • Author(s): Nakane T, Song C, Suzuki M, Nango E, Kobayashi J, Masuda T, Inoue S, Mizohata E, Nakatsu T, Tanaka T, Tanaka R, Shimamura T, Tono K, Joti Y, Kameshima T, Hatsui T, Yabashi M, Nureki O, Iwata S, Sugahara M.
  • Journal Title: Acta Crystallogr D Biol Crystallogr. Volume: 71 Issue: 12 Pages: 2519-25
  • DOI: 10.1107/s139900471501857x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structure and mechanism of the mammalian fructose transporter GLUT5 (2015)
  • Author(s): Nomura N, Verdon G, Kang HJ, Shimamura T, Nomura Y, Sonoda Y, Hussien SA, Qureshi AA, Coincon M, Sato Y, Abe H, Nakada-Nakura Y, Hino T, Arakawa T, Kusano-Arai O, Iwanari H, Murata T, Kobayashi T, Hamakubo T, Kasahara M, Iwata S, Drew D
  • Journal Title: Nature Volume: 526 Issue: 7573 Pages: 397-401
  • DOI: 10.1038/nature14909
  • NAID: 120005657713
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Grease matrix as a versatile carrier of proteins for serial crystallography. (2015)
  • Author(s): Sugahara M, Shimamura T, et al.
  • Journal Title: Nat Methods. Volume: 12 Issue: 1 Pages: 61-63
  • DOI: 10.1038/nmeth.3172
  • NAID: 120005682565
  • Peer Reviewed
• [Journal Article] Diverse application platform for hard X-ray diffraction in SACLA (DAPHNIS): application to serial protein crystallography using an X-ray free-electron laser. (2015)
  • Author(s): Tono K, Shimamura T, et al.
  • Journal Title: J. Synchrotron Rad. Volume: 22 Issue: 3 Pages: 532-537
  • DOI: 10.1107/s1600577515004464
  • Peer Reviewed
• [Journal Article] Proteoliposome-based selection of a recombinant antibody fragment against the human M2 muscarinic acetylcholine receptor. (2014)
  • Author(s): Suharni, Nomura Y, Arakawa T, Hino T, Abe H, Nakada-Nakura Y, Sato Y, Iwanari H, Shiroishi M, Asada H, Shimamura T, Murata T, Kobayashi T, Hamakubo T, Iwata S, Nomura N.
  • Journal Title: Monoclon Antib Immunodiagn Immunother Volume: 33 Issue: 6 Pages: 378-385
  • DOI: 10.1089/mab.2014.0041
  • NAID: 120005530933
  • Peer Reviewed
• [Journal Article] Crystal structure of a symmetric football-shaped GroEL:GroES2-ATP14 complex determined at 3.8Å reveals rearrangement between two GroEL rings. (2014)
  • Author(s): Koike-Takeshita A, Arakawa T, Taguchi H, Shimamura T.
  • Journal Title: J Mol Biol. Volume: 426 Issue: 21 Pages: 3634-41
  • DOI: 10.1016/j.jmb.2014.08.017
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] X線自由電子レーザーを用いたGPCRの構造解析 (2016)
  • Author(s): 島村達郎
  • Organizer: 第16回日本蛋白質科学会年会
  • Place of Presentation: 博多
  • Year and Date: 2016-06-08
  • Invited
• [Presentation] ヒスタミンH1受容体の構造解析とin silico screening (2014)
  • Author(s): 島村達郎
  • Organizer: 第87回日本内分泌学会学術総会
  • Place of Presentation: 博多
  • Year and Date: 2014-04-25
  • Invited
• [Book] Chapter 6 in Advanced Methods in Structural Biology (2016)
  • Author(s): Shimamura T.
  • Total Pages: 6
  • Publisher: Springer