Development of a method for simultaneous recordings of single channel dynamics and currents

• Project/Area Number: 15H04675
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General physiology
• Research Institution: University of Fukui
• Principal Investigator: Shimizu Hirofumi 福井大学, 学術研究院医学系部門, 講師 (50324158)
• Co-Investigator(Kenkyū-buntansha): 岩本 真幸 福井大学, 学術研究院医学系部門, 助教 (40452122) ; 平井 義和 京都大学, 工学(系)研究科(研究院), 研究員 (40452271)
• Research Collaborator: Tabata Osamu 京都大学, 大学院・工学系研究科, 教授 (20288624)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2017: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2016: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2015: ¥9,230,000 (Direct Cost: ¥7,100,000、Indirect Cost: ¥2,130,000)
• Keywords: 1分子計測 / 蛋白質 / X線回折 / イオンチャネル / タンパク質 / X線 / １分子計測(SMD) / マイクロ・ナノデバイス / Ｘ線

Outline of Final Research Achievements

Ion channel proteins are ion permeation pathway across cell membranes, whose functions are recorded with single-channel current recording method. The structure of KcsA potassium channel was first resolved at atomic resolutions among the ion channel proteins. We previously succeeded in recording conformational changes of the KcsA channel with the diffracted x-ray tracking method in a single-molecule level. In this study we tried to fabricate a new recording chamber and probes aiming to current-dynamics simultaneous recordings for the ion channel proteins.

Research Products

• [Journal Article] Concurrent In Vitro Synthesis and Functional Detection of Nascent Activity of the KcsA Channel under a Membrane Potential (2018)
  • Author(s): Iwamoto M, Elfaramawy MA, Yamatake M, Matsuura T, Oiki S
  • Journal Title: ACS Synthetic Biology Volume: 7 Issue: 4 Pages: 1004-1011
  • DOI: 10.1021/acssynbio.7b00454
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] 総説：MEMS研究と応用の最新動向 (2017)
  • Author(s): 平井義和
  • Journal Title: 表面技術 Volume: 68 Pages: 360-366
  • Peer Reviewed
• [Journal Article] Membrane Perfusion of Hydrophobic Substances Around Channels Embedded in the Contact Bubble Bilayer (2017)
  • Author(s): Iwamoto M, Oiki S
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 6857-6857
  • DOI: 10.1038/s41598-017-07048-4
  • Peer Reviewed / Open Access
• [Journal Article] Channel Formation and Membrane Deformation via Sterol-Aided Polymorphism of Amphidinol 3 (2017)
  • Author(s): Iwamoto M, Sumino A, Shimada E, Kinoshita M, Matsumori N, Oiki S
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 10782-10782
  • DOI: 10.1038/s41598-017-11135-x
  • Peer Reviewed / Open Access
• [Journal Article] Temperature-Jump effects on the single molecular dynamics of the KcsA Potassium Channels (2017)
  • Author(s): Hirofumi Shimizu, Masayuki Iwamoto
  • Journal Title: J. Physiol. Sci. Volume: 67 Suppl
• [Journal Article] “Effects of cholesterol on the function of the KcsA potassium channel” (2017)
  • Author(s): Masayuki Iwamoto, Shigetoshi Oiki
  • Journal Title: J. Physiol. Sci. Volume: 67 Suppl
• [Journal Article] Specific interactions between alkali metal cations and the KcsA channel studied using ATR-FTIR spectroscopy (2015)
  • Author(s): Y. Furutani, H. Shimizu, Y. Asai, S. Oiki, H. Kndori
  • Journal Title: Biophysics and Physicobiology Volume: 12 Issue: 0 Pages: 37-45
  • DOI: 10.2142/biophysico.12.0_37
  • NAID: 130005098800
  • ISSN: 2189-4779
  • Peer Reviewed / Open Access
• [Journal Article] Surface-enhanced IR absorption spectroscopy of the KcsA potassium channel upon application of an (2015)
  • Author(s): A. Yamakata, H. Shimizu, S. Oiki
  • Journal Title: Phys Chem Chem Phys. Volume: 17 Issue: 33 Pages: 21104-21111
  • DOI: 10.1039/c5cp02681d
  • Peer Reviewed / Open Access
• [Presentation] KcsAチャネル開閉構造変化の遷移過程における立体構造安定性地形 (2018)
  • Author(s): 清水啓史、岩本真幸
  • Organizer: 第95回日本生理学会
• [Presentation] 感光性基板接合材料を用いたX線1分子動態計測用マイクロ流体デバイスの作製 (2018)
  • Author(s): 森保彰, 田渕友樹, 平井義和, 清水啓史, 土屋智由, 田畑修
  • Organizer: 平成30年電気学会全国大会
• [Presentation] Development of a Method for Recording Single-Molecule Dynamics of Proteins in a Sub-millisecond time resolution (2018)
  • Author(s): Hirofumi Shimizu
  • Organizer: Pacific Rim Nano Medicine Symposium 2018 -The 9th Japan-Taiwan Symposium on Nanomedicine-
  • Int'l Joint Research
• [Presentation] KcsAカリウムイオンチャネルの1分子運動に対する試料温度ジャンプの影響 (2017)
  • Author(s): 清水啓史、岩本真幸
  • Organizer: 日本生理学会
  • Place of Presentation: アクトシティ浜松、浜松
  • Year and Date: 2017-03-28
• [Presentation] コレステロールがKcsAカリウムチャネル機能に及ぼす影響 (2017)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 日本生理学会
  • Place of Presentation: アクトシティ浜松、浜松
  • Year and Date: 2017-03-28
• [Presentation] Single-Molecule Fluctuations and Conformational Changes of KcsA Potassium Channels (2017)
  • Author(s): Hirofumi Shimizu
  • Organizer: The 8th Japan-Taiwan symposium on Nanomedicine
  • Place of Presentation: Academia Sinica, Taiwan
  • Year and Date: 2017-03-16
  • Int'l Joint Research / Invited
• [Presentation] 低X線散乱ノイズ観測チャンバー開発による蛋白質1分子構造変化計測 (2017)
  • Author(s): 清水啓史, 田渕友樹, 田原樹生, 岩本真幸, 平井義和, 土屋智由, 田畑修
  • Organizer: 第34回「センサ・マイクロマシンと応用システム」シンポジウム, 電気学会
• [Presentation] Structural Stabilities during Gating Transitions in KcsA Potassium Channel Revealed by Single-Molecule Dynamics Recordings (2017)
  • Author(s): Hirofumi Shimizu, Masayuki Iwamoto.,
  • Organizer: 第55回日本生物物理学会年会
• [Presentation] An Approach to Structural Analysis of a Small Membrane Protein KcsA by Cryo-electron Microscopy (2017)
  • Author(s): Hiroko Takazaki, Hirofumi Shimizu, Naoko Kajimura, Kaoru Mistuoka, Takuo Yasunaga
  • Organizer: 第55回日本生物物理学会年会
• [Presentation] ingle -Molecule Recordings of Gating Motions of KcsA Potassium Channels at Sub-millisecond Time Resolution (2017)
  • Author(s): Hirofumi Shimizu
  • Organizer: Conformational Ensembles from Experimental Data and Computer Simulations., Biophysical Society thematic meeting
  • Int'l Joint Research
• [Presentation] X 線1 分子動態計測法への試料温度ジャンプシステムの導入 (2016)
  • Author(s): 清水啓史、岩本真幸
  • Organizer: 日本生物物理学会
  • Place of Presentation: つくば国際会議場、つくば
  • Year and Date: 2016-11-25
• [Presentation] イオンチャネル機能に対する膜脂質効果の解析に向けた脂質二重膜組成の迅速変更法 (2016)
  • Author(s): 岩本真幸、老木成稔
  • Organizer: 日本生物物理学会
  • Place of Presentation: つくば国際会議場、つくば
  • Year and Date: 2016-11-25
• [Presentation] 生体膜を模倣した安定な人工脂質二重膜の作成 (2016)
  • Author(s): 老木成稔、岩本真幸
  • Organizer: バイオジャパン2016
  • Place of Presentation: パシフィコ横浜、横浜
  • Year and Date: 2016-10-12
• [Presentation] Photoresist Micro-Chamber for the Diffracted X-ray Tracking Method Recording Single-Molecule Conformational Changes (2016)
  • Author(s): Kio Tahara, Yoshikazu Hirai, Hirofumi Shimizu, Toshiyuki Tsuchiya, Osamu Tabata
  • Organizer: The 30th EUROSENSORS (EUROSENSORS XXX)
  • Place of Presentation: Budapest, Hungary
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] イオンチャネルの1分子構造変化計測 (2016)
  • Author(s): 清水啓史
  • Organizer: 第一回イオンチャネル研究会
  • Place of Presentation: 福岡大学、福岡
  • Year and Date: 2016-07-07
  • Invited
• [Presentation] KcsA チャネルゲーティングの膜脂質依存性 (2016)
  • Author(s): 岩本真幸
  • Organizer: 第一回イオンチャネル研究会
  • Place of Presentation: 福岡大学、福岡
  • Year and Date: 2016-07-07
• [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: 蛋白研セミナー
  • Place of Presentation: 大阪大学蛋白質研究所、吹田
• [Presentation] 蛋白質の分子揺らぎと構造変化を計測するX線１分子動態計測法の開発 (2015)
  • Author(s): 清水啓史
  • Organizer: 第53回日本生物物理学会年会、シンポジウム「動的構造生命科学を拓く新発想測定技術」（招待講演）
  • Place of Presentation: 金沢
  • Year and Date: 2015-09-13
• [Presentation] ３次元微細加工技術を応用したＸ線１分子動態計測用低ノイズマイクロチャンバの開発 (2015)
  • Author(s): 田原樹生,平井義和,清水啓史,土屋智由,田畑 修
  • Organizer: 電気学会総合研究会 E部門総合研究会
  • Place of Presentation: 福岡県 九州大学医学部百年講堂
  • Year and Date: 2015-07-02
• [Patent(Industrial Property Rights)] 脂質二重膜形成器 (2017)
  • Inventor(s): 老木成稔、岩本真幸
  • Industrial Property Rights Holder: 老木成稔、岩本真幸
  • Industrial Property Rights Type: 意匠
  • Filing Date: 2017
  • Acquisition Date: 2017
• [Patent(Industrial Property Rights)] 脂質平面膜を形成するための貫通孔を有するガラス基板、およびその製造方法と用途 (2016)
  • Inventor(s): 老木成稔、岩本真幸
  • Industrial Property Rights Holder: 老木成稔、岩本真幸
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016
  • Acquisition Date: 2017