Structural analyses of nucleic acid sensing TLR receptors

• Project/Area Number: 23770106
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: The University of Tokyo
• Principal Investigator: OHTO Umeharu 東京大学, 大学院・薬学系研究科, 助教 (90451856)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: タンパク質 / X線結晶構造解析 / 構造生物学 / 自然免疫 / Ｘ線結晶構造解析 / 結晶構造解析

Research Abstract

We conducted the protein preparation, crystallization, and crystal structural analyses on the nucleic acid sensing TLR receptor (TLR7, TLR8, TLR9). We succeeded the structural determination of the unliganded and liganded forms of TLR8. We obtained the sufficient amount of purified TLR7 and TLR9 for crystallization and obtained the crystal in several forms.

Research Products

• [Journal Article] Impaired α-TTP-PIPs Interaction Underlies Familial Vitamin E Deficiency. (2013)
  • Author(s): Kono, N., Ohto, U., Hiramatsu, T., Urabe, M., Uchida, Y., Satow, Y., and Arai, H.
  • Journal Title: Science Volume: 340 Pages: 1106-1110
  • NAID: 110009807290
  • Peer Reviewed
• [Journal Article] Structural reorganization of the Toll-like receptor8 dimer induced by agonistic ligands. (2013)
  • Author(s): Tanji H., Ohto U., Shibata T., Miyake M., and Shimizu T.
  • Journal Title: Science Volume: 339 Pages: 1426-1429
  • Peer Reviewed
• [Journal Article] TLR4-MD-2 complex is negatively regulated by an endogenous ligand, globotetraosylceramide. (2013)
  • Author(s): Kondo, Y., Ikeda, K., Tokuda, N.,Nishitani, C., Ohto, U., Akashi-Takamura, S., Ito, Y., Uchikawa, M., Kuroki, Y.,Taguchi, R., Miyake, K., Zhang, Q., Furukawa, K., and Furukawa, K.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 110 Pages: 4714-4719
  • Peer Reviewed
• [Journal Article] Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid (2013)
  • Author(s): Yamakawa N., Ohto U., Akashi-Takamura S., Takahashi K., Saitoh S., Tanimura N., Suganami T., Ogawa Y., Shibata T., Shimizu T., and Miyake K.
  • Journal Title: A. Int. Immunol. Volume: 25 Pages: 45-52
  • Peer Reviewed
• [Journal Article] Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid A (2013)
  • Author(s): N. Yamakawa, U. Ohto, S. Akashi-Takamura, K. Takahashi, SI. Saitoh, N. Tanimura, T. Suganami, Y. Ogawa, T. Shibata, T. Shimizu, K. Miyake
  • Journal Title: Int. Immunol. Volume: 25 Issue: 1 Pages: 45-52
  • DOI: 10.1093/intimm/dxs084
  • Peer Reviewed
• [Journal Article] TLR4-MD-2 complex is negatively regulated by an endogenous ligand, globotetraosylceramide in vascular endothelial cells. (2013)
  • Author(s): Kondo, Y., Tokuda, N., Nishitani, C., Ohto, U., Akashi-TakKondo, Y., Tokuda, N., Nishitani, C., Ohto, U., Akashi-Takamura, S., Ito, Y., Uchikawa, M., Kuroki, Y., Miyake, K., Zhang, Q., Furukawa, K., Furukawa, K
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 110 Issue: 12 Pages: 4714-4719
  • DOI: 10.1073/pnas.1218508110
  • Peer Reviewed
• [Journal Article] Structural reorganization of the Toll-like receptor 8 dimer induced by agonistic ligands (2013)
  • Author(s): Tanji H, Ohto U, Shibata T, Miyake K, Shimizu T
  • Journal Title: Science Volume: 339 Issue: 6126 Pages: 1426-1429
  • DOI: 10.1126/science.1229159
  • Peer Reviewed
• [Journal Article] Structural analyses of human TLR4 polymorphism D299G and T399I. (2012)
  • Author(s): Ohto, U., Yamakawa, N., Akashi-Takamura, S., Miyake, K., & Shimizu, T.
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 40611-40617
  • Peer Reviewed
• [Journal Article] Coagulation factor X activates innate immunity to human species C adenovirus. (2012)
  • Author(s): Doronin, K., Flatt, J. W., Di Paolo, N. C., Khare, R., Kalyuzhniy, O., Acchione, M.,Sumida, J. P., Ohto, U., Shimizu, T., Akashi-Takamura, S., Miyake, K., MacDonald, J. W., Mammler, T. K., Beyer, R. P., Farin, F. M., Stewart, P. L., Shayakhmetov, D. M.
  • Journal Title: Science Volume: 338 Pages: 795-798
  • Peer Reviewed
• [Journal Article] Structural basis of species-specific endotoxin sensing by TLR4/MD-2. (2012)
  • Author(s): Ohto, U., Fukase, K., Miyake, K., & Shimizu, T.
  • Journal Title: Proc. Natl. Acad. Sci. USA. Volume: 109 Pages: 7421-7426
  • Peer Reviewed
• [Journal Article] Saturated Fatty Acid and TLR Signaling Link βCell Dysfunction and Islet Inflammation. (2012)
  • Author(s): Eguchi, K., Manabe, I., Oishi-Tanaka, Y., Ohsugi, M., Kono, N., Ogata, F., Yagi,N., Ohto, U., Kimoto, M., Miyake, K., Tobe, K., Arai, H., Kadowaki, T., & Nagai, R.
  • Journal Title: Cell Metab. Volume: 15 Pages: 1-16
  • Peer Reviewed
• [Journal Article] Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human β-galactosidase. (2012)
  • Author(s): Usui, K., Ohto, U., Ochi, T., Shimizu, T.,& Satow, Y.
  • Journal Title: Acta Crystallogr. Sect. Volume: F68 Pages: 73-77
  • Peer Reviewed
• [Journal Article] Crystal Structure of Human β-Galactosidase: STRUCTURAL BASIS OF GM1GANGLIOSIDOSIS AND MORQUIO B DISEASES. (2012)
  • Author(s): Ohto, U., Usui, K., Ochi, T., Yuki, K., Satow, Y., & Shimizu, T.
  • Journal Title: J. Biol. Chem. Volume: 287 Pages: 1801-1812
  • Peer Reviewed
• [Journal Article] Coagulation Factor X Activates Innate Immunity to Human Species C Adenovirus. (2012)
  • Author(s): Doronin K.
  • Journal Title: Science. Volume: 6108 Issue: 6108 Pages: 795-8
  • DOI: 10.1126/science.1226625
  • Peer Reviewed
• [Journal Article] Saturated fatty acid and TLR signaling link β cell dysfunction and islet inflammation. (2012)
  • Author(s): Eguchi K, Manabe I, Oishi-Tanaka Y, Ohsugi M, Kono N, Ogata F, Yagi N, Ohto U, Kimoto M, Miyake K, Tobe K, Arai H, Kadowaki T, Nagai R
  • Journal Title: Cell Metab. Volume: 15 Issue: 4 Pages: 518-533
  • DOI: 10.1016/j.cmet.2012.01.023
  • Peer Reviewed
• [Journal Article] Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human β-galactosidase (2012)
  • Author(s): Usui, K., Ohto, U., Ochi, T., Shimizu, T., Satow, Y.
  • Journal Title: Acta Crystallogr.Sect.F Volume: 68 Issue: 1 Pages: 73-77
  • DOI: 10.1107/s1744309111047920
  • Peer Reviewed
• [Journal Article] Crystal Structure of Human β-Galactosidase: The Structural Basis of GM1 Gangliosidosis and Morquio B Diseases (2012)
  • Author(s): Ohto U, et al.
  • Journal Title: J.Biol.Chem. Volume: 287 Issue: 3 Pages: 1801-1812
  • DOI: 10.1074/jbc.m111.293795
  • Peer Reviewed
• [Journal Article] Crystal structures of mouse and human RP105/MD-1 complexes reveal unique dimer organization of the toll-like receptor family. (2011)
  • Author(s): Ohto, U., Miyake, K., & Shimizu, T.
  • Journal Title: J. Mol. Biol. Volume: 413 Pages: 815-825
  • Peer Reviewed
• [Presentation] 自然免疫系RNA認識受容体TLR8の結晶構造 (2013)
  • Author(s): 大戸梅治、丹治裕美、柴田琢磨、三宅健介、清水敏之
  • Organizer: 第22回東京免疫フォーラム
  • Place of Presentation: 東京
  • Year and Date: 2013-03-14
• [Presentation] 病原体由来RNAを認識する自然免疫系TLR8の結晶構造 (2013)
  • Author(s): 大戸梅治,丹治裕美,柴田琢磨,三宅健介,清水敏之
  • Organizer: 日本薬学会第133年会
  • Place of Presentation: 横浜
• [Presentation] Structural basis of species-specific endotoxin sensing by TLR4/MD-2 (2012)
  • Author(s): Umeharu Ohto
  • Organizer: Program for Young Researcher Symposium in Europe
  • Place of Presentation: Cambridge, UK
  • Year and Date: 2012-06-07
• [Presentation] 自然免疫系RP105/MD-1複合体の構造解析 (2012)
  • Author(s): 大戸梅治,三宅健介,清水敏之
  • Organizer: 日本薬学会132年会
  • Place of Presentation: 札幌
  • Year and Date: 2012-03-29
• [Presentation] 4本鎖lipidAによる種特異的TLR4/MD-2活性化機構の構造基盤 (2012)
  • Author(s): 大戸梅治,深瀬浩一,三宅健介,清水敏之
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 博多
• [Presentation] 4本鎖lipid Aによる種特異的TLR4/MD-2活性化機構の構造基盤 (2012)
  • Author(s): 大戸 梅治, 深瀬浩一，三宅健介，清水敏之
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 博多
• [Presentation] Structural analyses of mouse MD-1 protein complexed with endogenous phospholipid (2011)
  • Author(s): Umeharu Ohto, Hitomi Harada, and Yoshinori Satow
  • Organizer: International Union of Crystallography 2011
  • Place of Presentation: Madrid, Spain
• [Remarks]
  • URL: http://www.f.u-tokyo.ac.jp/~kouzou/index.html
• [Remarks]
  • URL: http://www.f.u-tokyo.ac.jp/~kouzou/index.html