Structural and functional analysis of gap junction channels using electron microscopy

• Project/Area Number: 26840019
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: Nagoya University
• Principal Investigator: Oshima Atsunori 名古屋大学, 名古屋大学細胞生理学研究センター, 准教授 (80456847)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: ギャップ結合チャネル / イネキシン / 電子顕微鏡 / 三次元再構成 / 二次元結晶 / 極低温電子顕微鏡 / 二次元結晶化

Outline of Final Research Achievements

In this project, the structure of C. elegans innexin-6 (INX-6) was investigated using electron crystallography. Two dimensional crystals of N-terminal deleted INX-6 where 2 to 19 were eliminated were successfully obtained. The structure of INX-6delN reveals the six-teen subunits, a hexadecamer. The plug density was observed on the channel pore pathway, suggesting a structure in a closed form. These results have been published as a research article

Research Products

• [Journal Article] Hexadecameric structure of an invertebrate gap junction channel (2016)
  • Author(s): Oshima, A., Matsuzawa, T., Murata, K., Tani, K. and Fujiyoshi, Y.
  • Journal Title: Journal of Molecular Biology Volume: 428 Issue: 6 Pages: 1227-1236
  • DOI: 10.1016/j.jmb.2016.02.011
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] ギャップ結合チャネルの構造と多様性 (2016)
  • Author(s): 大嶋篤典
  • Journal Title: 膜 Volume: 41 Pages: 50-56
  • NAID: 130005249079
  • Peer Reviewed
• [Journal Article] GraDeR: membrane protein complex preparation for single particle cryo-EM. (2015)
  • Author(s): Hauer F., Gerle C., Fischer N., Oshima A., Shinzawa-Itoh K., Shimada S., Yokoyama K., Fujiyoshi Y. & Stark H.
  • Journal Title: Structure Volume: 23 Issue: 9 Pages: 1769-1775
  • DOI: 10.1016/j.str.2015.06.029
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] THREE-DIMENSIONAL STRUCTURE OF INNEXIN GAP JUNCTION CHANNELS STUDIED BY ELECTRON CRYSTALLOGRAPHY (2016)
  • Author(s): Atsunori Oshima, Tomohiro Matsuzawa, Kazuyoshi Murata, Kazutoshi Tani and Yoshinori Fujiyoshi
  • Organizer: 60th Biophysical Society Annual Meeting
  • Place of Presentation: Los Angeles Convention Center, Los Angeles
  • Year and Date: 2016-02-27
  • Int'l Joint Research
• [Presentation] Three dimensional structure of C. elegans innexin gap junction channels (2015)
  • Author(s): Atsunori Oshima, Tomohiro Matsuzawa, Kazuyoshi Murata, Kazutoshi Tani and Yoshinori Fujiyoshi
  • Organizer: 第38回日本分子生物学会年会、第88回日本生化学会大会 合同大会
  • Place of Presentation: 神戸ポートアイランド、神戸
  • Year and Date: 2015-12-01
• [Presentation] Initial 3D reconstruction of innexin gap junction channels by electron crystallography (2015)
  • Author(s): Atsunori Oshima,, Tomohiro Matsuzawa, Kazuyoshi Murata, Kazutoshi Tani and Yoshinori Fujiyoshi
  • Organizer: 日本顕微鏡学会 第71回学術講演会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2015-05-13 – 2015-05-15
• [Presentation] Initial 3D reconstruction of innexin gap junction channels by electron crystallography (2015)
  • Author(s): Atsunori Oshima,, Tomohiro Matsuzawa, Kazuyoshi Murata, Kazutoshi Tani and Yoshinori Fujiyoshi
  • Organizer: 日本顕微鏡学会 第71回学術講演会
  • Place of Presentation: 国立京都国際会館、京都
  • Year and Date: 2015-05-13
• [Presentation] Structure and diversity of gap junction channels studied by electron microscopy (2015)
  • Author(s): Atsunori Oshima
  • Organizer: 第120回日本解剖学会総会・全国学術集会・第92回日本生理学会大会合同大会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2015-03-21 – 2015-03-23
  • Invited
• [Presentation] Electron Microscopic Studies of Invertebrate Gap Junction Channels (2015)
  • Author(s): Atsunori Oshima, Tomohiro Matsuzawa, Kazuyoshi Murata, Kouki Nishikawa, Kazutoshi Tani and Yoshinori Fujiyoshi,
  • Organizer: IGER International Symposium on Frontiers in Biological Research with Advanced Electron Microscope Technologies
  • Place of Presentation: Nagoya University
  • Year and Date: 2015-01-15 – 2015-01-16
  • Invited
• [Presentation] 電子顕微鏡で見るギャップ結合チャネルの構造と多様性 (2014)
  • Author(s): 大嶋篤典
  • Organizer: 「電子顕微鏡機能イメージングの医学・生物学への応用」～次世代の生物電顕を考える～
  • Place of Presentation: 生理学研究所
  • Year and Date: 2014-11-12 – 2014-11-13
  • Invited
• [Presentation] Electron microscopy of C. elegans innexin-6 gap junction channels indicates the characteristic subunit organization (2014)
  • Author(s): Atsunori Oshima, Tomohiro Matsuzawa, Kazuyoshi Murata, Kouki Nishikawa, and Yoshinori Fujiyoshi
  • Organizer: 第52回日本生物物理学会年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2014-09-25 – 2014-09-27
  • Invited
• [Presentation] Oligomeric form and function of C. elegans innexin-6 gap junction channels (2014)
  • Author(s): Atsunori Oshima, Tomohiro Matsuzawa, Kazuyoshi Murata, Kouki Nishikawa, and Yoshinori Fujiyoshi
  • Organizer: the 23rd International Union of Crystallography (IUCr) Congress and General Assembly
  • Place of Presentation: Montréal, Québec, Canada
  • Year and Date: 2014-08-05 – 2014-08-12
  • Invited
• [Presentation] 電子顕微鏡によるイネキシンギャップ結合チャネルの構造研究 (2014)
  • Author(s): 大嶋篤典、松澤朋寛、村田和義、西川幸希、藤吉好則
  • Organizer: 日本顕微鏡学会第70回記念学術講演会
  • Place of Presentation: 幕張メッセ 国際会議場
  • Year and Date: 2014-05-11 – 2014-05-13