Development and application of the experimental platform for the single-molecule analysis of the lipid-effects on the ion channel function

• Project/Area Number: 25440067
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: University of Fukui
• Principal Investigator: Iwamoto Masayuki 福井大学, 医学部, 助教 (40452122)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: 生体膜 / 1分子計測 / チャネル / 脂質 / 電気生理 / １分子計測

Outline of Final Research Achievements

Based on the principle of formation of the contact bilayer between water-in-oil droplets, we developed the platform for the artificial membrane experiments to analyze quantitatively the "lipid-effects" on the function of the membrane proteins. We prepared a variety of lipid bilayer membranes having different lipid composition including asymmetric membranes on the platform. We reconstituted the KcsA potassium channel into these membranes and succeeded in detecting the lipid-dependency in the activity of the channel at single-molecule level.

Research Products

• [Journal Article] Rectified Proton Grotthuss Conduction Across a Long Water-Wire in the Test Nanotube of the Polytheonamide B Channel. (2016)
  • Author(s): Matsuki, Y., Iwamoto, M. Mita, K., Shigemi, K., Matsunaga, S., and Oiki
  • Journal Title: J. Am. Chem. Soc. Volume: 138 Issue: 12 Pages: 4168-4177
  • DOI: 10.1021/jacs.5b13377
  • NAID: 120006226473
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Mechanism for attenuated outward conductance induced by mutations in the cytoplasmic pore of Kir2.1 channels. (2015)
  • Author(s): Chang, H.-K., Iwamoto, M., Oiki, S. and Shieh, R.-C.
  • Journal Title: Sci. Rep. Volume: 5 Issue: 1 Pages: 18404-18404
  • DOI: 10.1038/srep18404
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Methods for the functional analysis of ion channels on the contact bubble bilayer by the intra-bubble perfusion (2015)
  • Author(s): Iwamoto M, Oiki S
  • Journal Title: The Journal of Physiological Sciences Volume: 65 Suppl.
• [Journal Article] Contact bubble bilayers with flush drainage (2015)
  • Author(s): Iwamoto M., Oiki S.
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 9110-9110
  • DOI: 10.1038/srep09110
  • NAID: 120005572953
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] pH-dependent promotion of phospholipid flip-flop by the KcsA potassium channel (2015)
  • Author(s): H. Nakao, K. Ikeda, M. Iwamoto, H. Shimizu, S. Oiki, Y. Ishihama, M. Nakano
  • Journal Title: Biochim. Biophys. Acta-Biomembr. Volume: 1848 Issue: 1 Pages: 145-150
  • DOI: 10.1016/j.bbamem.2014.10.001
  • Peer Reviewed
• [Journal Article] Paradoxical one-ion pore behavior of the long β-helical peptide of marine cytotoxic polytheonamide B (2014)
  • Author(s): M. Iwamoto, S. Matsunaga, S. Oiki
  • Journal Title: Sci. Rep. Volume: 4 Issue: 1 Pages: 3636-3636
  • DOI: 10.1038/srep03636
  • Peer Reviewed / Open Access
• [Journal Article] Gating-associated clustering-dispersion dynamics of the KcsA potassium channel in a lipid membrane (2014)
  • Author(s): A. Sumino, D. Yamamoto, M. Iwamoto, T. Dewa, S. Oiki
  • Journal Title: J. Phys. Chem. Lett. Volume: 5 Issue: 3 Pages: 578-584
  • DOI: 10.1021/jz402491t
  • Peer Reviewed
• [Presentation] 膜脂質がカリウムイオンチャネルのゲート開閉を制御する分子機構 (2016)
  • Author(s): 岩本真幸
  • Organizer: 大阪大学蛋白質研究所セミナー「膜タンパク質の構造ダイナミクス」
  • Place of Presentation: 大阪大学蛋白質研究所（大阪）
  • Year and Date: 2016-05-13
  • Invited
• [Presentation] 油中水滴接触膜でのイオンチャネル電流測定中に脂質二重膜組成を瞬間的に変更する試み (2016)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 第93回日本生理学会大会
  • Place of Presentation: 札幌コンベンションセンター（札幌）
  • Year and Date: 2016-03-23
• [Presentation] 液滴接触膜を応用したイオンチャネル機能解析法 (2015)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 第62回中部日本生理学会
  • Place of Presentation: 富山大学五福キャンパス（富山）
  • Year and Date: 2015-11-13
• [Presentation] 流動電位測定により明らかにされたカリウムチャネルポア内でのイオンと水の共役的透過 (2015)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 日本生物物理学会第53回年会（シンポジウム「電気生理学的アプローチによる膜タンパク質構造機能相関」）
  • Place of Presentation: 金沢大学角間キャンパス（金沢）
  • Year and Date: 2015-09-15
  • Invited
• [Presentation] 微小な液滴接触膜の形成とイオンチャネル機能解析への応用 (2015)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 日本生物物理学会第53回年会
  • Place of Presentation: 金沢大学角間キャンパス（金沢）
  • Year and Date: 2015-09-14
• [Presentation] 液滴内潅流による液滴接触膜上でのチャネル機能解析法 (2015)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 第92回日本生理学会大会
  • Place of Presentation: 神戸コンベンションセンター（神戸）
  • Year and Date: 2015-03-21
• [Presentation] Molecular mechanism of the membrane lipid-dependent gating of the KcsA potassium channel (2014)
  • Author(s): Iwamoto M., Oiki S.
  • Organizer: 第45回生理研国際シンポジウム
  • Place of Presentation: 岡崎コンファレンスセンター（岡崎）
  • Year and Date: 2014-11-26 – 2014-11-28
• [Presentation] 脂質はイオンチャネルのゲート開閉をどの様に制御するのか？：新奇脂質センサーによる制御機 (2014)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（札幌）
  • Year and Date: 2014-09-26
  • Invited
• [Presentation] Modulation of the activation gating of the KcsA potassium channel by the membrane lipids (2014)
  • Author(s): Iwamoto M, Oiki S
  • Organizer: Single Protein Dynamics in Cellulo 2014
  • Place of Presentation: 沖縄科学技術大学院大学（沖縄県恩納村）
  • Year and Date: 2014-04-21 – 2014-04-25
• [Presentation] Effect of membrane lipids on the inactivation gating of the KcsA potassium channel (2014)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 第91回日本生理学会大会
  • Place of Presentation: 鹿児島大学（鹿児島市）
• [Presentation] Membrane lipids modulate the inactivation gating of the KcsA potassium channel (2014)
  • Author(s): Iwamoto M., Oiki S.
  • Organizer: Biophysical Society 58th Annual Meeting
  • Place of Presentation: Moscone Center (San Francisco, Califolnia, USA)
• [Presentation] Rolling of N-terminal amphipathic helix on the anionic inner membrane leaflet stabilizes the open state of the KcsA potassium channel (2013)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 日本生物物理学会第51回年会
  • Place of Presentation: 京都国際会議場（京都市）
• [Remarks] 分子生理学（生理学１）研究室
  • URL: http://seiri1.med.lab.u-fukui.ac.jp/
• [Remarks] 分子生理学（生理学１）研究室
  • URL: http://seiri1.med.lab.u-fukui.ac.jp/ja/