Study on channel protein gating dynamics by single molecule measurement.

• Project/Area Number: 22370059
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biophysics
• Research Institution: Okayama University (2013-2014); The Graduate School for the Creation of New Photonics Industries (2011-2012); The Institute of Physical and Chemical Research (2010)
• Principal Investigator: IDE TORU 岡山大学, 自然科学研究科, 教授 (60231148)
• Project Period (FY): 2010-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥19,110,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥4,410,000); Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2011: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2010: ¥9,750,000 (Direct Cost: ¥7,500,000、Indirect Cost: ¥2,250,000)
• Keywords: １分子計測(SMD) / 生物物理学 / イオンチャネル / １分子計測 / 単一チャネル電流 / 人工膜 / 蛍光 / 1分子計測

Outline of Final Research Achievements

In order for elucidating the structure-function relationship of ion-channel proteins, we have developed an apparatus for simultaneous optical and electrical measurement of single ion-channel proteins. Using this apparatus, we successfully measured single ligand bindings to single ion-channel proteins. We developed a novel method for measuring ion-channel current in the course of apparatus development process, which is applicable to ion-channel sensor devices. We revealed a relationship between the structure and the function of potassium channel, KcsA, using macroscopic methods.

Research Products

• [Journal Article] A single amino acid gates the KcsA channel. (2014)
  • Author(s): Hirano, M, Okuno, D, Onishi, Y, Ide, T.
  • Journal Title: Biochem Biophys Res Commun. Volume: 450(4) Issue: 4 Pages: 1537-1540
  • DOI: 10.1016/j.bbrc.2014.07.032
  • Peer Reviewed
• [Journal Article] Automated Parallel Recordings of Topologically Identified Single Ion Channels. (2013)
  • Author(s): Kawano R, Tsuji Y, Sato K, Osaki T, Kamiya K, Hirano M, Ide T, Miki N, Takeuchi S.
  • Journal Title: Scientific Reports Volume: 3 Issue: 1 Pages: 1955-1955
  • DOI: 10.1038/srep01995
  • Peer Reviewed / Open Access
• [Journal Article] Uncovering the Protein Translocon at the Chloroplast Inner Envelope Membrane (2013)
  • Author(s): Shingo Kikuchi
  • Journal Title: SCIENCE Volume: 339 Issue: 6119 Pages: 571-574
  • DOI: 10.1126/science.1229262
  • Peer Reviewed
• [Journal Article] Lab on a Chip (2012)
  • Author(s): Y. Oyama
  • Journal Title: A glass fiber sheet-based electroosmotic lateral flow immunoassay for point of care testing. Volume: 12 Issue: 24 Pages: 5155-5159
  • DOI: 10.1039/c2lc40994a
  • Peer Reviewed
• [Journal Article] 「ちから」でイオンチャネルの機能を制御する (2012)
  • Author(s): 井出徹
  • Journal Title: 生物物理 Volume: 52 Pages: 289-290
  • NAID: 10031126103
• [Journal Article] Direct Manipulation of a Single Potassium Channel Gate with an Atomic Force Microscope Probe (2011)
  • Author(s): M.Kitta, T.Ide, M.Hirano, H.Tanaka, T.Yanagida, T.Kawai
  • Journal Title: Small Volume: 7(16) Issue: 16 Pages: 2379-2383
  • DOI: 10.1002/smll.201002337
  • Peer Reviewed
• [Journal Article] The role of the KcsA channel cytoplasimic domain in pH-dependent gating (2011)
  • Author(s): M.Hirano, Y.Onishi, T.Yanagida, T.Ide
  • Journal Title: Biophysical Journal Volume: 101(9) Issue: 9 Pages: 2157-2162
  • DOI: 10.1016/j.bpj.2011.09.024
  • Peer Reviewed
• [Journal Article] Channels formed by amphotericin B covalent dimers exhibit rectification (2011)
  • Author(s): M.Hirano, Y.Takeuchi, N.Matsumori, M.Murata, T.Ide
  • Journal Title: J Membr Biol Volume: 240 Issue: 3 Pages: 159-164
  • DOI: 10.1007/s00232-011-9354-x
  • Peer Reviewed
• [Journal Article] Rearrangements in the KcsA cytoplasmic domain underlie its gating. (2010)
  • Author(s): Hirano Minako
  • Journal Title: J Biol Chem Volume: 285 Pages: 3777-3783
  • Peer Reviewed
• [Journal Article] Single channel properties of lysenin measured in artificial lipid bilayers and their applications to biomolecule detection. (2010)
  • Author(s): Takaaki Aoki
  • Journal Title: Proc Jpn Acad Ser B Phys Biol Sci Volume: 86 Pages: 920-925
  • Peer Reviewed
• [Journal Article] Simultaneous optical and electrical recording of single drug bonding to single ion channel proteins. (2010)
  • Author(s): Ide T.
  • Journal Title: ChemPhysChem Volume: 11 Pages: 3408-3411
  • Peer Reviewed
• [Presentation] A simple method for lipid bilayer formation using a fine gold electrode. (2015)
  • Author(s): Daichi Okuno, Minako Hirano, Hiroaki Yokota, Tomotaka Komori, Yukiko Onishi, Toshio Yanagida, Toru Ide
  • Organizer: The Joint symposium of 9th International Symposium on Medical, Bio- and Nano-Electronics, and 6th International Workshop on Nanostructures & Nanoelectronics
  • Place of Presentation: 東北大学
  • Year and Date: 2015-03-02
  • Invited
• [Presentation] Simple method for lipid bilayer formation with simultaneous incorporation of ion channels using gold electrode. (2014)
  • Author(s): Daichi Okuno, Minako Hirano, Hiroaki Yokota, Tomotaka Komori, Yukiko Onishi, Toshio Yanagida, Toru Ide
  • Organizer: The 52nd Annual Meeting of the Biophysical Society of Japan
  • 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] Modifications of ion channel function. The 52nd Annual Meeting of the Biophysical Society of Japan (2014)
  • Author(s): Minako Hirano, Daichi Okuno, Yukiko Onishi, Hiroaki Yokota, Toru Ide
  • Organizer: The 52nd Annual Meeting of the Biophysical Society of Japan
  • 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 the line confocal system for the single molecule tracking of fast folding dynamics of proteins. (2014)
  • Author(s): Hiroyuki Oikawa, Kiyoto Kamagata, Munehito Arai, Atsuhito Fukasawa, Hiroaki Yokota, Toru Ide, Satoshi Takahashi
  • Organizer: The 52nd Annual Meeting of the Biophysical Society of Japan
  • 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] The KcsA channel cytoplasmic domain effects on the inactivation gating (2013)
  • Author(s): M. Hirano
  • Organizer: Biophysical Society 57th Annual Meeting
  • Place of Presentation: Philadelphia
• [Presentation] チャネルタンパクの1分子計測 (2012)
  • Author(s): 井出徹
  • Organizer: JSTさきがけ「ナノテク融合」領域懇話会
  • Place of Presentation: 金沢市(石川県)
  • Year and Date: 2012-01-13
• [Presentation] Reconstitution of ion channel into lipid bilayer using glass needle. (2012)
  • Author(s): D. Okuno
  • Organizer: The 50th annual meeting of the biophysical society of Japan
  • Place of Presentation: Nagoya Univ.
• [Presentation] Single molecule imaging of bio-molecules. (2012)
  • Author(s): Toru Ide
  • Organizer: The 14th Takayanagi Kenjiro Memorial Symposium
  • Place of Presentation: Shizuoka Univ.
  • Invited
• [Presentation] The KcsA channel cytoplasmic domain effects on the inactivation gating (2012)
  • Author(s): M. Hirano
  • Organizer: The 50th annual meeting of the biophysical society of Japan
  • Place of Presentation: Nagoya Univ.
• [Presentation] チャネルタンパクの1分子計測 (2011)
  • Author(s): 井出徹
  • Organizer: 遺伝研バイオロジカルシンポジウム
  • Place of Presentation: 遺伝研(静岡県)
  • Year and Date: 2011-12-07
• [Presentation] Mechanism for opening and closing by the KcsA channel (2011)
  • Author(s): M.Hirano, Y.Onishi, T.Yanagida, T.Ide
  • Organizer: 第49回日本生物物理学会
  • Place of Presentation: 兵庫県立大学(兵庫県)
  • Year and Date: 2011-09-18
• [Presentation] Direct Manipulation of a Single Potassium Channel Gate with an Atomic Force Microscope Probe (2011)
  • Author(s): T.Ide, M.Hirano, D.Okuno, M.Kitta
  • Organizer: 第49回日本生物物理学会
  • Place of Presentation: 兵庫県立大学(兵庫県)
  • Year and Date: 2011-09-18
• [Presentation] The KcsA channel opening and closing mechanism (2011)
  • Author(s): Minako Hirano
  • Organizer: QBiC Retreat 2011 : How to achieve "Whole Cell Modeling"
  • Place of Presentation: ゆにとぴあ篠山(兵庫県)
  • Year and Date: 2011-09-10
• [Presentation] イオンチャネルタンパクの1分子計測・操作法の開発 (2011)
  • Author(s): 井出徹
  • Organizer: 静岡大学ナノバイオ科学講演会
  • Place of Presentation: 静岡大学(静岡県)
  • Year and Date: 2011-07-28
• [Presentation] 蛍光色素を利用したイオンチャネルの開閉機構の可視化 (2011)
  • Author(s): 平野美奈子
  • Organizer: 新学術領域研究「細胞機能と分子活性の多次元蛍光生体イメージング」第1回研究代表者会議
  • Place of Presentation: 北海道大学(北海道)
  • Year and Date: 2011-06-30
• [Presentation] The role of the cytoplasmic domain in pH-dependent gating by the Kcs A channel (2011)
  • Author(s): Minako Hirano
  • Organizer: Biophysical Society 55th Annual Meeting
  • Place of Presentation: Baltimore, Maryland US
  • Year and Date: 2011-03-07
• [Presentation] The role of the cytoplasmic domain in pH-dependent gating by the KcsA channel (2010)
  • Author(s): Minako Hirano
  • Organizer: Molecular Science of Fluctuations toward Biological Functions, The 4rd International Symposium
  • Place of Presentation: 滋賀県大津市
  • Year and Date: 2010-12-01
• [Presentation] The role of the cytoplasmic domain in pH-dependent gating by the Kcs A channel (2010)
  • Author(s): Minako Hirano
  • Organizer: GCOE/Structural Biology Research Center International Symposium
  • Place of Presentation: 愛知県名古屋市
  • Year and Date: 2010-11-23
• [Presentation] The role of the cytoplasmic domain in pH-dependent gating by the Kcs A channel (2010)
  • Author(s): 平野美奈子
  • Organizer: 第48回 日本生物物理学会年会
  • Place of Presentation: 宮城県仙台市
  • Year and Date: 2010-09-22
• [Presentation] 可視1分子計測によるチャネルたんぱく質の分子内運動計測 (2010)
  • Author(s): 井出徹
  • Organizer: 第30回表面科学学術講演会・第51回真空に関する連合講演会合同講演会
  • Place of Presentation: 大阪府吹田市
  • Year and Date: 2010-09-04
• [Presentation] Kチャネルゲートの1分子操作 (2010)
  • Author(s): 井出徹
  • Organizer: 第10回日本蛋白質科学会年会 蛋白質1分子内部運動と機能発現
  • Place of Presentation: 北海道札幌市
  • Year and Date: 2010-06-17
• [Presentation] Coordination between the cytoplasmic domain and the inactivation gate in the KcsA channel
  • Author(s): M. Hirano, Y. Onishi, D. Okuno, T. Ide
  • Organizer: 第51回 日本生物物理学会
  • Place of Presentation: 国立京都国際会館
• [Presentation] Reconstitution of ion channel immobilized on solid support into lipid bilayer
  • Author(s): D. Okuno, M. Hirano, Y. Onishi, T. Ide
  • Organizer: 第51回 日本生物物理学会
  • Place of Presentation: 国立京都国際会館
• [Presentation] 膜タンパクの１分子計測・操作
  • Author(s): 井出徹
  • Organizer: 高分子学会・バイオ高分子研究会
  • Place of Presentation: 蒲郡
  • Invited
• [Book] 細胞工学8月号 “明らかになった葉緑体のトランスロコン” (2013)
  • Author(s): 菊池 真吾, 平野 美奈子, 井出 徹, 中井 正人
  • Total Pages: 2
  • Publisher: 学研メディカル秀潤社
• [Book] Cell Signaling Reactions : Single-Molecular Kinetic Analysis(Simultaneous optical and electrical recording of single drug bindings to single ion channel proteins.)(Ed.by Y.Sako and M.Ueda) (2011)
  • Author(s): Toru Ide
  • Total Pages: 14
  • Publisher: Springer