一回膜貫通型受容体のシグナル伝達の構造基盤

• Project Area: Integrative understanding of biological signaling networks based on mathematical science
• Project/Area Number: 17H05991
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Complex systems
• Research Institution: The University of Tokyo
• Principal Investigator: 大戸 梅治 東京大学, 大学院薬学系研究科(薬学部), 准教授 (90451856)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000); Fiscal Year 2018: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000); Fiscal Year 2017: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
• Keywords: シグナル伝達 / 一回膜貫通型受容体 / 構造生物学

Outline of Annual Research Achievements

生体中において一回膜貫通型受容体は種々の局面で重要な役割を果たしている。例えば免疫において微生物など病原体の構成成分は一回膜貫通型受容体であるToll-like receptors (TLRs)などの病原体センサーによって認識され様々な免疫応答を引き起こす。これら一回膜貫通型受容体は、様々な病気の治療薬のターゲットとされている。しかし、一回膜貫通型受容体の機能発現機構は主に細胞外のシグナル受容ドメインと細胞内のシグナル伝達ドメイン個別に研究されてきた。そのため、細胞外ドメインと細胞内ドメインの間で細胞膜を介してどのような協働性が存在してシグナルが伝達されるのかに関する実験的証拠があまりにも乏しいため、現状では前者と後者の間の関連性はほとんど憶測の域を出ない。本課題では，主に全長の受容体を用いてこの細胞外ドメインと細胞内ドメインの協働性を構造生物学的に明らかにし、真の意味でのリガンド認識とシグナル伝達機構を明らかにすることを目的とする。
上記を達成するために、今年度はTLR全長の電顕解析に向けた試料調製に取り組んだ。特にTLR3全長に関して、ナノディスクに再構成する条件の検討を詳細に行った。その結果、１つのナノディスクに対して１分子のTLR3全長を再構成することに成功した。また再構成した試料について、ネガティブ染色電顕観察とクライオ電顕観察を進めた。また、再構成した試料についてリガンドが結合することを確認している。今後、さらに検討を進め、TLR全長の構造解析を目指す。

Research Progress Status

平成30年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

平成30年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Toll-like Receptor 9 Contains Two DNA Binding Sites that Function Cooperatively to Promote Receptor Dimerization and Activation (2018)
  • Author(s): Ohto Umeharu、Ishida Hanako、Shibata Takuma、Sato Ryota、Miyake Kensuke、Shimizu Toshiyuki
  • Journal Title: Immunity Volume: 48 Issue: 4 Pages: 649-658
  • DOI: 10.1016/j.immuni.2018.03.013
  • Peer Reviewed / Open Access
• [Journal Article] Structural basis for species-specific activation of mouse Toll-like receptor 9 (2018)
  • Author(s): Ishida Hanako、Ohto Umeharu、Shibata Takuma、Miyake Kensuke、Shimizu Toshiyuki
  • Journal Title: FEBS Letters Volume: 592 Issue: 15 Pages: 2636-2646
  • DOI: 10.1002/1873-3468.13176
  • Peer Reviewed / Open Access
• [Journal Article] Small-Molecule TLR8 Antagonists via Structure-Based Rational Design (2018)
  • Author(s): Hu Zhenyi、Tanji Hiromi、Jiang Shuangshuang、Zhang Shuting、Koo Kyoin、Chan Jean、Sakaniwa Kentaro、Ohto Umeharu、Candia Albert、Shimizu Toshiyuki、Yin Hang
  • Journal Title: Cell Chemical Biology Volume: 25 Issue: 10 Pages: 1286-1291
  • DOI: 10.1016/j.chembiol.2018.07.004
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Structural analysis of HTL and D14 proteins reveals the basis for ligand selectivity in Striga (2018)
  • Author(s): Xu Yuqun、Miyakawa Takuya、Nosaki Shohei、Nakamura Akira、Lyu Ying、Nakamura Hidemitsu、Ohto Umeharu、Ishida Hanako、Shimizu Toshiyuki、Asami Tadao、Tanokura Masaru
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 3947-3947
  • DOI: 10.1038/s41467-018-06452-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structural Analyses of Toll-like Receptor 7 Reveal Detailed RNA Sequence Specificity and Recognition Mechanism of Agonistic Ligands (2018)
  • Author(s): Zhang Zhikuan、Ohto Umeharu、Shibata Takuma、Taoka Masato、Yamauchi Yoshio、Sato Ryota、Shukla Nikunj M.、David Sunil A.、Isobe Toshiaki、Miyake Kensuke、Shimizu Toshiyuki
  • Journal Title: Cell Reports Volume: 25 Issue: 12 Pages: 3371-3381
  • DOI: 10.1016/j.celrep.2018.11.081
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mechanisms controlling nucleic acid-sensing Toll-like receptors (2018)
  • Author(s): Miyake Kensuke、Shibata Takuma、Ohto Umeharu、Shimizu Toshiyuki、Saitoh Shin-Ichiroh、Fukui Ryutaro、Murakami Yusuke
  • Journal Title: International Immunology Volume: 30 Issue: 2 Pages: 43-51
  • DOI: 10.1093/intimm/dxy016
  • Peer Reviewed / Open Access
• [Journal Article] Small-molecule inhibition of TLR8 through stabilization of its resting state (2018)
  • Author(s): Zhang, S., Hu, Z., Tanji, H., Jiang, S., Das, N, Li, J., Sakaniwa, K., Jin, J., Bian, Y., Ohto, U., Shimizu, T., Yin, H.
  • Journal Title: Nature Chem. Biol. Volume: 14 Issue: 1 Pages: 58-64
  • DOI: 10.1038/nchembio.2518
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The crystal structure of the AhRR-ARNT heterodimer reveals the structural basis of the repression of AhR-mediated transcription (2017)
  • Author(s): Sakurai, S., Shimizu, T., & *Ohto, U.
  • Journal Title: J. Biol. Chem. Volume: 292 Issue: 43 Pages: 17609-17616
  • DOI: 10.1074/jbc.m117.812974
  • Peer Reviewed
• [Journal Article] Funiculosin variants and phosphorylated derivatives promote innate immune responses via the Toll-like receptor 4/myeloid differentiation factor-2 complex (2017)
  • Author(s): Okamoto N, Mizote K, Honda H, Saeki A, Watanabe Y, Yamaguchi-Miyamoto T, Fukui R, Tanimura N, Motoi Y, Akashi-Takamura S, Kato T, Fujishita S, Kimura T, Ohto U, Shimizu T, Hirokawa T, Miyake K, Fukase K, Fujimoto Y, Nagai Y, Takatsu K
  • Journal Title: Journal of Biological Chemistry Volume: 292 Issue: 37 Pages: 15378-15394
  • DOI: 10.1074/jbc.m117.791780
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Towards a structural understanding of nucleic acid-sensing Toll-like receptors in the innate immune system (2017)
  • Author(s): Zhang, Z., Ohto, U., & Shimizu, T.
  • Journal Title: FEBS letters. Volume: 591 Issue: 20 Pages: 3167-3181
  • DOI: 10.1002/1873-3468.12749
  • Peer Reviewed
• [Journal Article] Conservation and Divergence of Ligand Recognition and Signal Transduction Mechanisms in Toll-Like Receptors (2017)
  • Author(s): Ohto, U.
  • Journal Title: Chemical and Pharmaceutical Bulletin Volume: 65 Issue: 8 Pages: 697-705
  • DOI: 10.1248/cpb.c17-00323
  • NAID: 130005875868
  • ISSN: 0009-2363, 1347-5223
  • Peer Reviewed
• [Journal Article] The crystal structure of human DEAH-box RNA helicase 15 reveals a domain organization of the mammalian DEAH/RHA family. (2017)
  • Author(s): Murakami, K., Nakano, K., Shimizu, T., & *Ohto, U.
  • Journal Title: Acta Crystallogr F Struct Biol Commun. Volume: 73 Issue: 6 Pages: 347-355
  • DOI: 10.1107/s2053230x17007336
  • Peer Reviewed
• [Presentation] 自然免疫を担う病原体センサー蛋白質の構造基盤の解明と創薬展開 (2019)
  • Author(s): 大戸梅治、清水敏之
  • Organizer: 日本薬学会 第139年会
  • Invited
• [Presentation] The simultaneous binding of two DNA motifs for efficient activation of TLR9 revealed by X-ray Crystallography (2018)
  • Author(s): Umeharu Ohto, Toshiyuki Shimizu
  • Organizer: Toll meeting 2018
  • Int'l Joint Research
• [Presentation] LIGAND RECOGNITION MECHANISM OF TLR7 REVEALED BY A LARGE-SCALE CRYSTALLOGRAPHIC STUDY (2018)
  • Author(s): Z. Zhang, U. Ohto, T.Shibata, R.Sato, K.Miyake, T. Shimizu
  • Organizer: Toll meeting 2018
  • Int'l Joint Research
• [Presentation] Structural biology of Toll-like receptors (2018)
  • Author(s): Umeharu Ohto, Toshiyuki Shimizu
  • Organizer: 第56回日本生物物理学会
  • Invited
• [Presentation] 自然免疫受容体TLR9活性化機構の構造生物学的研究 (2018)
  • Author(s): 石田英子，大戸梅治，柴田琢磨，佐藤亮太，三宅健介，清水敏之
  • Organizer: 日本結晶学会平成30年度年会
• [Presentation] AhRR-ARNTヘテロダイマーによるAhRの転写抑制機構の構造基盤 (2018)
  • Author(s): 櫻井駿也、清水敏之、大戸梅治
  • Organizer: 2017年度量子ビームサイエンスフェスタ
• [Presentation] AhRR-ARNTヘテロダイマーによるAhRの転写抑制機構の構造基盤 (2018)
  • Author(s): 櫻井駿也、清水敏之、大戸梅治
  • Organizer: 日本薬学会 第138年会
• [Presentation] Sperm-egg interaction revealed by the structure of IZUMO1-JUNO complex (2017)
  • Author(s): Umeharu Ohto
  • Organizer: American Society for Reproductive Medicine (ASRM) 2017 Scientific Congress
  • Int'l Joint Research / Invited
• [Presentation] Structural analysis reveals ligand recognition mechanism of Toll-like receptor 7 (2017)
  • Author(s): Zhikuan Zhang, Umeharu Ohto, Toshiyuki Shimizu
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] sor (AhRR)によるAhRの転写抑制機構の構造基盤解明 (2017)
  • Author(s): 櫻井駿也、清水敏之、大戸梅治
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] Aryl hydrocarbon receptorの機能発現機構解明に向けた構造生物学的研究 (2017)
  • Author(s): 櫻井駿也、大戸梅治、清水敏之
  • Organizer: 第17回日本蛋白質科学会年会
• [Presentation] The crystal structure of AhRR/ARNT complex (2017)
  • Author(s): Shunya Sakurai, Umeharu Ohto, Toshiyuki Shimizu
  • Organizer: 24th Congress and General Assembly of the International Union of Crystallography
  • Int'l Joint Research
• [Presentation] 自然免疫Toll様受容体のリガンド認識および活性化機構 (2017)
  • Author(s): 大戸梅治
  • Organizer: 第61回日本薬学会関東支部大会
  • Invited
• [Presentation] Crystal structure of the autoinhibited form of NOD2 (2017)
  • Author(s): Umeharu Ohto, Sakiko Maekawa, Toshiyuki Shimizu
  • Organizer: The 67th Annual Meeting of the American Crystallographic Association (ACA 2017)
  • Int'l Joint Research
• [Presentation] Strcutrual studies of Toll-like receptor 7 (2017)
  • Author(s): Zhikuan Zhang, Umeharu Ohto, Toshiyuki Shimizu
  • Organizer: The 67th Annual Meeting of the American Crystallographic Association (ACA 2017)
  • Int'l Joint Research
• [Remarks] 研究室ホームページ
  • URL: http://www.f.u-tokyo.ac.jp/~kouzou/index.html