Structural biology of thermosensation in human

• Project/Area Number: 25711004
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Partial Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Tottori University
• Principal Investigator: Hino Tomoya 鳥取大学, 工学(系)研究科(研究院), 准教授 (40373360)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥27,040,000 (Direct Cost: ¥20,800,000、Indirect Cost: ¥6,240,000); Fiscal Year 2015: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥7,020,000 (Direct Cost: ¥5,400,000、Indirect Cost: ¥1,620,000); Fiscal Year 2013: ¥14,560,000 (Direct Cost: ¥11,200,000、Indirect Cost: ¥3,360,000)
• Keywords: TRPチャネル / イオンチャネル / 膜タンパク質 / X線結晶構造解析 / タンパク質精製 / タンパク質発現 / 酵母 / 立体構造解析 / 温度感知 / 構造生物学 / 温度受容 / 機能性抗体

Outline of Final Research Achievements

Thermosensation is an essential physiological process for the proper adaptation to the changes in environmental temperature. In this study, to gain insight into the molecular mechanisms of thermosensation operated by TRP ion channels, we challenged to determine the atomic structures of human TRP channels.
We successfully overexpressed 9 thermosensitive TRP channels　by employing methylotrophic yeast Pichia pastoris as a host strain and purified 5 channels by utilizing affinity column that is immobilized GFP-nanobody. In addition, we obtained several monoclonal antibodies that recognize conformational epitopes of TRPV4.

Research Products

• [Journal Article] Crystal structure of D-stereospecific amidohydrolase from Streptomyces sp. 82F2: insight into the structural factors for substrate specificity. (2016)
  • Author(s): Arima J, Shimone K, Miyatani K, Tsunehara Y, Isoda Y, Hino T, Nagano S
  • Journal Title: FEBS Journal Volume: 283 Issue: 2 Pages: 337-349
  • DOI: 10.1111/febs.13579
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Crystal structure of the anion exchanger domain of human erythrocyte band 3 (2015)
  • Author(s): Arakawa T, Kobayashi-Yurugi T, Alguel Y, Iwanari H, Hatae H, Iwata M, Abe Y, Hino T, Ikeda-Suno C, Kuma H, Kang D, Murata T, Hamakubo T, Cameron AD, Kobayashi T, Hamasaki N, Iwata S
  • Journal Title: Science Volume: 350 Issue: 6261 Pages: 680-684
  • DOI: 10.1126/science.aaa4335
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The crystal structure of heme acquisition system A from Yersinia pseudotuberculosis (HasA ypt): Roles of the axial ligand Tyr75 and two distal arginines in heme binding (2015)
  • Author(s): Kanadani M., Sato T., Hino T., Nagano S., Ozaki S.
  • Journal Title: Journal of inorganic biochemistry Volume: 151 Pages: 26-33
  • DOI: 10.1016/j.jinorgbio.2015.07.007
  • Peer Reviewed
• [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] Crystal structures of the human adiponectin receptors. (2015)
  • Author(s): Tanabe H, Fujii Y, Okada-Iwabu M, Iwabu M, Nakamura Y, Hosaka T, Motoyama K, Ikeda M, Wakiyama M, Terada T, Ohsawa N, Hato M, Ogasawara S, Hino T, Murata T, Iwata S, Hirata K, Kawano Y, Yamamoto M, Kimura-Someya T, Shirouzu M, Yamauchi T, Kadowaki T, Yokoyama S
  • Journal Title: Nature Volume: 520 Issue: 7547 Pages: 312-316
  • DOI: 10.1038/nature14301
  • NAID: 40020952688
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] ヒト由来TRPM8の立体構造解析に向けた高純度精製試料調製の検討 (2015)
  • Author(s): 川越 雄斗，落合 晶也，三牧 裕哉，日野 智也，永野 真吾
  • Organizer: 第88回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-02