Direct visualization of functional basis in voltage-dependent ion-channels

• Project/Area Number: 18K06172
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43040:Biophysics-related
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Shigematsu Hideki 国立研究開発法人理化学研究所, 放射光科学研究センター, 研究員 (00415928)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2019: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2018: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: クライオ電子顕微鏡 / 構造生物学 / イオンチャネル / 電位依存性 / 膜電位 / 可視化 / 構造・機能相関

Outline of Final Research Achievements

Conformational change in ion-pore has been revealed directly by using single particle electron cryo-microscopy in this study. The specimen used for image analysis had been utilized in functional analysis by using florescent spectroscopy and it revealed that it can maintain membrane potential in a few minutes. The method here can be applied to any other membrane proteins to see its function on close to physiological environment, liposome.

Academic Significance and Societal Importance of the Research Achievements

リポソーム上に提示された膜タンパク質の機能構造をクライオ電子顕微鏡で明らかにすることは、今後の医薬品開発への応用や、実際に細胞が機能する現場を捉えるという基礎学問的な意義も重要である。

Research Products

• [Int'l Joint Research] Yale University(米国)
• [Int'l Joint Research] Yale University(米国)
• [Int'l Joint Research] Yale University(米国)
• [Journal Article] Cryo-EM Structure of the Prostaglandin E Receptor EP4 Coupled to G Protein. (2021)
  • Author(s): Nojima S, Fujita Y, Kimura KT, Nomura N, Suno R, Morimoto K, Yamamoto M, Noda T, Iwata S, Shigematsu H, Kobayashi T.
  • Journal Title: Structure Volume: 29 Issue: 3 Pages: 252-260
  • DOI: 10.1016/j.str.2020.11.007
  • Peer Reviewed / Open Access
• [Journal Article] Cryo-EM structures of Toll-like receptors in complex with UNC93B1 (2021)
  • Author(s): Ishida Hanako、Asami Jinta、Zhang Zhikuan、Nishizawa Tomohiro、Shigematsu Hideki、Ohto Umeharu、Shimizu Toshiyuki
  • Journal Title: Nature Structural & Molecular Biology Volume: 28 Issue: 2 Pages: 173-180
  • DOI: 10.1038/s41594-020-00542-w
  • Peer Reviewed / Open Access
• [Journal Article] GTP-dependent formation of straight tubulin oligomers leads to microtubule nucleation (2021)
  • Author(s): Ayukawa Rie、Iwata Seigo、Imai Hiroshi、Kamimura Shinji、Hayashi Masahito、Ngo Kien Xuan、Minoura Itsushi、Uchimura Seiichi、Makino Tsukasa、Shirouzu Mikako、Shigematsu Hideki、Sekimoto Ken、Gigant Benoit、Muto Etsuko
  • Journal Title: Journal of Cell Biology Volume: 220 Issue: 4 Pages: 1-19
  • DOI: 10.1083/jcb.202007033
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Electron cryo-microscopy for elucidating the dynamic nature of live-protein complexes (2020)
  • Author(s): Shigematsu Hideki
  • Journal Title: Biochimica et Biophysica Acta (BBA) - General Subjects Volume: 1864 Issue: 2 Pages: 129436-129436
  • DOI: 10.1016/j.bbagen.2019.129436
  • Peer Reviewed
• [Journal Article] Structural analysis reveals TLR7 dynamics underlying antagonism (2020)
  • Author(s): Tojo Shingo、Zhang Zhikuan、Matsui Hiroyuki、Tahara Masahiro、Ikeguchi Mitsunori、Kochi Mami、Kamada Mami、Shigematsu Hideki、Tsutsumi Akihisa、Adachi Naruhiko、Shibata Takuma、Yamamoto Masaki、Kikkawa Masahide、Senda Toshiya、Isobe Yoshiaki、Ohto Umeharu、Shimizu Toshiyuki
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 1-11
  • DOI: 10.1038/s41467-020-19025-z
  • Peer Reviewed / Open Access
• [Journal Article] Local structural changes of the influenza A virus ribonucleoprotein complex by single mutations in the specific residues involved in efficient genome packaging. (2019)
  • Author(s): Takizawa N, Ogura Y, Fujita Y, Noda T, Shigematsu H, Hayashi T, Kurokawa K
  • Journal Title: Virology Volume: 531 Pages: 126-140
  • DOI: 10.1016/j.virol.2019.03.004
  • Peer Reviewed / Open Access
• [Journal Article] HCV IRES captures an actively translating 80S ribosome (2019)
  • Author(s): Yokoyama Takeshi、Machida Kodai、Iwasaki Wakana、Shigeta Tomoaki、Nishimoto Madoka、Takahashi Mari、Sakamoto Ayako、Yonemochi Mayumi、Harada Yoshie、Shigematsu Hideki、Shirouzu Mikako、Tadakuma Hisashi、Imataka Hiroaki、Ito Takuhiro
  • Journal Title: Molecular Cell Volume: 印刷中 Issue: 6 Pages: 1205-1214
  • DOI: 10.1016/j.molcel.2019.04.022
  • Peer Reviewed / Open Access
• [Journal Article] Kinesin-binding-triggered conformation switching of microtubules contributes to polarized transport (2018)
  • Author(s): Shima Tomohiro、Morikawa Manatsu、Kaneshiro Junichi、Kambara Taketoshi、Kamimura Shinji、Yagi Toshiki、Iwamoto Hiroyuki、Uemura Sotaro、Shigematsu Hideki、Shirouzu Mikako、Ichimura Taro、Watanabe Tomonobu M.、Nitta Ryo、Okada Yasushi、Hirokawa Nobutaka
  • Journal Title: The Journal of Cell Biology Volume: 217 Issue: 12 Pages: 4164-4183
  • DOI: 10.1083/jcb.201711178
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Structural insight into microtubule stabilization and kinesin inhibition by Tau family MAPs (2018)
  • Author(s): Shigematsu Hideki、Imasaki Tsuyoshi、Doki Chihiro、Sumi Takuya、Aoki Mari、Uchikubo-Kamo Tomomi、Sakamoto Ayako、Tokuraku Kiyotaka、Shirouzu Mikako、Nitta Ryo
  • Journal Title: The Journal of Cell Biology Volume: 217 Issue: 12 Pages: 4155-4163
  • DOI: 10.1083/jcb.201711182
  • NAID: 120006599259
  • Peer Reviewed
• [Journal Article] Structural basis of epilepsy-related ligand-receptor complex LGI1-ADAM22 (2018)
  • Author(s): Yamagata Atsushi、Miyazaki Yuri、Yokoi Norihiko、Shigematsu Hideki、Sato Yusuke、Goto-Ito Sakurako、Maeda Asami、Goto Teppei、Sanbo Makoto、Hirabayashi Masumi、Shirouzu Mikako、Fukata Yuko、Fukata Masaki、Fukai Shuya
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 1546-1546
  • DOI: 10.1038/s41467-018-03947-w
  • Peer Reviewed / Open Access
• [Presentation] Progress in Cryo-EM Imaging of Kv1.2 Channels with Membrane Potential Applied (2019)
  • Author(s): Hideki Shigematsu, Youshan Yang, Yangyang Yan, Fred J. Sigworth
  • Organizer: Gordon Research Conference
  • Int'l Joint Research
• [Presentation] Spring-8における共用を目指したクライオ 電子顕微鏡施設 (2019)
  • Author(s): 重松秀樹
  • Organizer: 日本顕微鏡学会第75回学術講演会
• [Presentation] Cryo-EM Imaging of Kv1.2 Channels with Membrane Potential Applied (2019)
  • Author(s): Hideki Shigematsu, Youshan Yang, Yangyang Yan, Fred J. Sigworth
  • Organizer: Biophysical Society Annual Meetings
  • Int'l Joint Research
• [Presentation] 高分解能クライオ電子顕微鏡単粒子解析の検討 (2018)
  • Author(s): 重松秀樹
  • Organizer: 日本顕微鏡学会第74回学術講演会
• [Presentation] クライオ電顕による電位依存性チャネルの作動原理の可視化 (2018)
  • Author(s): Hideki Shigematsu, Youshan Yang, Yanyang Yan, Fred Sigworth
  • Organizer: 第41回日本分子生物学会年会