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

• Project/Area Number: 24249013
• Research Category: Grant-in-Aid for Scientific Research (A)
• 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, 医学部, 講師 (50324158)
• Co-Investigator(Renkei-kenkyūsha): IWAMOTO Masayuki 福井大学, 医学部, 助教 (40452122)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥47,710,000 (Direct Cost: ¥36,700,000、Indirect Cost: ¥11,010,000); Fiscal Year 2014: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2013: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2012: ¥37,180,000 (Direct Cost: ¥28,600,000、Indirect Cost: ¥8,580,000)
• Keywords: 蛋白質 / 1分子動態計測 / 1分子計測(SMD) / 生理学 / １分子計測（SMD） / X線

Outline of Final Research Achievements

Prior to realizing simultaneous recordings of single-molecular dynamics and currents we tried to increase time-resolution of the dynamics measurementin this project. We intorduced a X-ray focusing mirror and a spectrum measurement system to the sychrotorn facility(SPring8). The time resolutions of the dynamics measurements became to be in a sub-millisecond level, which is equivalent to that of single channel current measurement. And the X-ray radiation damages of the ion channel proteins were decreased by using the spectrum measurement system. These advances of the method will lead to the development of simultaneous measurement system in near future.

Research Products

• [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] Structural Changes of the KcsA Potassium Channel Upon Application of the Electrode Potential Studied by Surface-Enhanced IR Absorption Spectroscopy (2013)
  • Author(s): Akira Yamakata, Hirofumi Shimizu, Masatoshi Osawa, Shigetoshi Oiki
  • Journal Title: Chemical Physics Volume: 419 Pages: 224-228
  • DOI: 10.1016/j.chemphys.2013.02.035
  • Peer Reviewed
• [Journal Article] ATR-FTIR Spectroscopy Revealed the Different Vibrational Modes of the Selectivity Filter Interacting with K+ and Na+ in the Open and Collapsed Conformations of the KcsA Potassium Channel (2012)
  • Author(s): Furutani, Y., Shimizu, H., Asai, Y., Fukuda, T., Oiki, S. and Kandori, H
  • Journal Title: J. Phys. Chem. Lett Volume: 3 Issue: 24 Pages: 3806-3810
  • DOI: 10.1021/jz301721f
  • Peer Reviewed
• [Presentation] Relationship between Structural Fluctuations and The Opening Conformational Changes of The KcsA (2015)
  • Author(s): Hirofumi Shimizu, Masayuki Iwamoto, Yumiko Oota, Shigetoshi Oiki
  • 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
  • Place of Presentation: 神戸国際会議場・展示場
  • Year and Date: 2015-03-21 – 2015-03-23
• [Presentation] The diffracted X-ray tracking method for recording the single-molecule conformational changes of the KcsA potassium channel in a sub-millisecond time resolution (2014)
  • Author(s): Hirofumi Shimizu, Masayuki Iwamoto, Yumiko Oota, Yoshikazu Hirai, Osamu Tabata, Shigetoshi Oiki
  • Organizer: The 45th Natl Inst Physiol Sci (NIPS) International Symposium
  • Place of Presentation: Okazaki, Japan
  • Year and Date: 2014-11-25 – 2014-11-28
• [Presentation] Ｘ線１分子動態計測法で用いる低ノイズ観測チャンバーの開発 (2014)
  • Author(s): 平井義和、田畑修、清水啓史
  • Organizer: 化学とマイクロ・ナノシステム学会 第30回研究会（CHEMINAS30）
  • Place of Presentation: 北海道大学フロンティア応用化学研究棟
  • Year and Date: 2014-10-02 – 2014-10-03
• [Presentation] The Enhancement of Structural Fluctuations Prior to The Opening Conformational Changes of The (2014)
  • Author(s): Hirofumi Shimizu, Masayuki Iwamoto, Yumiko Oota, Shigetoshi Oiki
  • Organizer: 第52回 日本生物物理学会
  • 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
• [Presentation] Development of a method for measuring single-molecule motions of the KcsA potassium channel in a (2014)
  • Author(s): Hirofumi Shimizu, Masayuki Iwamoto, Antoine Royant , David Von Stetten,
  • Organizer: Single Protein Dynamics in Cellulo 2014: Spatio-Temporal, Structural
  • Place of Presentation: OIST seaside house, Okinawa
  • Year and Date: 2014-04-21 – 2014-04-25
• [Presentation] Development of a method for measuring single-molecule motions of the KcsA potassium channel in a sub-millisecond time resolution (2014)
  • Author(s): Hirofumi Shimizu , Masayuki Iwamoto , Antoine Royant, David Von Stetten, Laurent Guerin, Michael Wulff , Shigetoshi Oiki
  • Organizer: Single Protein Dynamics in Cellulo 2014:Spatio-Temporal, Structural and Quantitative Analyses
  • Place of Presentation: 沖縄、OISTシーサイドハウス
• [Presentation] 高時間分解能で蛋白質の分子揺らぎと構造変化を計測するためのX線1分子動態計測法の開発 (2013)
  • Author(s): 清水啓史、岩本真幸、老木成稔
  • Organizer: 日本生物物理学会
  • Place of Presentation: 京都、国立京都国際会館
• [Presentation] イオンチャネル開閉構造変化のX線1分子動画計測 ─分子揺らぎと構造変化の相関の解明を目指して─ (2013)
  • Author(s): 清水啓史
  • Organizer: 日本物理学会第68回年次大会 シンポジウム「タンパク質の動きを解き明かす一分子計測の最前線」
  • Place of Presentation: 広島
  • Invited
• [Presentation] Development of a method for recording single molecular gating dynamics of KcsA potassium channel in a sub-millisecond time resolution (2013)
  • Author(s): Hirofumi Shimizu
  • Organizer: Nagoya Symposium Frontiers in Structural Physiology
  • Place of Presentation: 名古屋
• [Presentation] KcsA カリウムイオンチャネルの溶液条件変化応答1 分子開閉ダイナミクスの解析 (2012)
  • Author(s): 清水啓史
  • Organizer: 第50回日本生物物理学会年会，シンポジウム「先端顕微計測が照らす生命の輝き」
  • Place of Presentation: 名古屋
  • Invited
• [Presentation] カリウムチャネルの選択透過性とゲーティング (2012)
  • Author(s): 老木成稔，岩本真幸，炭竃享司，清水啓史
  • Organizer: 平成24年度生理学研究所研究会「膜機能分子の機能・構造ゆらぎの時空間スペクトル解析」
  • Place of Presentation: 岡崎
• [Presentation] サブミリ秒時間スケールでのKcsA カリウムイオンチャネル開閉構造変化の1 分子動画計測 (2012)
  • Author(s): 清水啓史
  • Organizer: 第12回日本蛋白質科学会年会，シンポジウム「YouTube 時代の構造生物学」
  • Place of Presentation: 名古屋
  • Invited
• [Remarks] イオンチャネル蛋白質1分子の動きを動画でとらえる ～X線1分子動態計測法の開発～
  • URL: https://nanonet.go.jp/magazine/