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Single molecule measurement of ligand bindings to the acetylcholine receptor channel

Research Project

Project/Area Number 18K06157
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43040:Biophysics-related
Research InstitutionOkayama University

Principal Investigator

Ide Toru  岡山大学, ヘルスシステム統合科学学域, 教授 (60231148)

Project Period (FY) 2018-04-01 – 2023-03-31
Project Status Completed (Fiscal Year 2022)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywordsイオンチャネル / 1分子計測 / 単一チャネル電流計測 / 1分子イメージング / 1分子計測 / チャネル / アセチルコリン受容体
Outline of Final Research Achievements

Single molecule imaging system utilizing the local surface plasmon at the tip of a sharp gold electrode was developed. The electrode was fabricated by electro-chemical etching the tip of a thin gold wire. An automatic etching system was developed to increase fabrication efficiency. Fluorescence observation was performed using fluorescent dyes with low quantum yields, and single molecules of fluorescent dyes were observed.
The electrode surface was modified to be hydrophilic, and an artificial lipid bilayer membrane was successfully formed on the electrode surface. Using this artificial membrane, single channel current analysis of channel proteins, channel-forming peptides, and pore-forming toxins was successfully performed.
The above artificial membrane method has succeeded in significantly improving the efficiency of channel current measurement compared to conventional methods, and has also enabled automated measurement using a robot.

Academic Significance and Societal Importance of the Research Achievements

チャネル蛋白は機能の1分子解析(単一チャネル電流解析)が最も進んだ蛋白である。各々のチャネルに対して、詳細な状態遷移モデルが提案されており、本研究の成果はモデルの直接検証手段を与える。また、チャネルは1分子のリガンド結合と機能の相関を高時間分解で観測できる蛋白であり、計測によって得られた知見を蛋白一般の分子機構の理解に拡げることも可能となる。また、チャネル蛋白は膜のイオンの透過性を制御することよって、多岐に渡る生命活動おいて重要な役割を果たしている。チャネル蛋白一般の活性化分子機構を詳細に知ることは、科学的な興味に止まらず、医療、創薬などの分野においても重要である。

Report

(6 results)
  • 2022 Annual Research Report   Final Research Report ( PDF )
  • 2021 Research-status Report
  • 2020 Research-status Report
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (21 results)

All 2023 2022 2021 2020 2019 2018

All Journal Article (8 results) (of which Peer Reviewed: 8 results,  Open Access: 5 results) Presentation (13 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Simple Methods for Single Ion-channel Recordings2023

    • Author(s)
      平野 美奈子,朝倉 真実,井出 徹
    • Journal Title

      Seibutsu Butsuri

      Volume: 63 Issue: 2 Pages: 110-114

    • DOI

      10.2142/biophys.63.110

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2022 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Kastor and Polluks polypeptides encoded by a single gene locus cooperatively regulate VDAC and spermatogenesis.2022

    • Author(s)
      Mise S#, Matsumoto A#*, Shimada K, Hosaka T, Takahashi M, Ichihara K, Shimizu H, Shiraishi C, Saito D, Suyama M, Yasuda T, Ide T, Izumi Y, Bamba T, Kimura-Someya T, Shirouzu M, Miyata H, Ikawa M, Nakayama KI*.
    • Journal Title

      Nat. Commun.

      Volume: 13(1) Issue: 1 Pages: 1071-1071

    • DOI

      10.1038/s41467-022-28677-y

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Development of an automated system to measure ion channel currents using a surface-modified gold probe.2021

    • Author(s)
      Minako Hirano, Masahisa Tomita, Chikako Takahashi, Nobuyuki Kawashima and Toru Ide
    • Journal Title

      Scientific reports

      Volume: 11(1) Issue: 1 Pages: 17934-17934

    • DOI

      10.1038/s41598-021-97237-z

    • Related Report
      2021 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Characteristics of channel pores formed by Bacillus thuringiensis mosquito-larvicidal Cry4Aa toxin2021

    • Author(s)
      Shiraishi Yuri、Shiozaki Tomoya、Asakura Mami、Ide Toru、Hayakawa Tohru
    • Journal Title

      Applied Entomology and Zoology

      Volume: 57 Issue: 1 Pages: 63-70

    • DOI

      10.1007/s13355-021-00762-6

    • Related Report
      2021 Research-status Report
    • Peer Reviewed
  • [Journal Article] Channel-pore cation selectivity is a major determinant of Bacillus thuringiensis Cry46Ab mosquitocidal activity2020

    • Author(s)
      Hayakawa Tohru、Miyazaki Midoka、Harada Syoya、Asakura Mami、Ide Toru
    • Journal Title

      Applied Microbiology and Biotechnology

      Volume: 104 Issue: 20 Pages: 8789-8799

    • DOI

      10.1007/s00253-020-10893-5

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] A Lipid Bilayer Formed on a Hydrogel Bead for Single Ion Channel Recordings2020

    • Author(s)
      Hirano Minako、Yamamoto Daiki、Asakura Mami、Hayakawa Tohru、Mise Shintaro、Matsumoto Akinobu、Ide Toru
    • Journal Title

      Micromachines

      Volume: 11 Issue: 12 Pages: 1070-1070

    • DOI

      10.3390/mi11121070

    • Related Report
      2020 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Characterization of the channel-pores formed by Bacillus thuringiensis Cry46Ab toxin in planar lipid bilayers.2019

    • Author(s)
      Sakakibara A, Takebe S, Ide T, Hayakawa T.
    • Journal Title

      Applied Entomology and Zoology

      Volume: 54 Issue: 4 Pages: 389-398

    • DOI

      10.1007/s13355-019-00635-z

    • NAID

      120006877709

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Journal Article] Electrostatic state of the cytoplasmic domain influences inactivation at the selectivity filter of the KcsA potassium channel.2019

    • Author(s)
      Minako Hirano, Toru Ide
    • Journal Title

      Biochimica et Biophysica Acta (BBA) - Biomembranes

      Volume: 1861(1) Issue: 1 Pages: 220-227

    • DOI

      10.1016/j.bbamem.2018.07.011

    • Related Report
      2018 Research-status Report
    • Peer Reviewed
  • [Presentation] An artificial lipid bilayer ion-channel recording method using agarose gel beads2022

    • Author(s)
      Mami Asakura, Atsuya Mukuno, Minako Hirano, Toru Ide
    • Organizer
      The 60th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2022 Annual Research Report
  • [Presentation] Artificial bilayers on a hydrogel bead for channel current recordings2021

    • Author(s)
      Daiki Yamamoto, Mami Asakura, Minako Hirano, Toru Ide
    • Organizer
      The 59th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2021 Research-status Report
  • [Presentation] Development of a novel channel current measurement method using agarose gel beads2021

    • Author(s)
      Mami Asakura, Daiki Yamamoto, Minako Hirano, Toru Ide
    • Organizer
      The 59th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2021 Research-status Report
  • [Presentation] Development of an automated system for measuring channel currents using a gold probe2021

    • Author(s)
      Minako Hirano, Masahisa Tomita, Chikako Takahashi, Nobuyuki Kawashima, Toru Ide
    • Organizer
      The 59th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2021 Research-status Report
  • [Presentation] Channel-pores formation of Bacillus thuringiensis Cry46Ab toxin and its mosquitocidal activity2021

    • Author(s)
      Midoka Miyazaki, Tohru Hayakawa, Toru Ide
    • Organizer
      The 59th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2021 Research-status Report
  • [Presentation] Development of a system for automated ionic current measurement2020

    • Author(s)
      Minako Hirano, Masahisa Tomita , Chikako Takahashi, Nobuyuki Kawashima, Toru Ide
    • Organizer
      The 58th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2020 Research-status Report
    • Int'l Joint Research
  • [Presentation] Artificial bilayers on hydrogel for channel current recordings2020

    • Author(s)
      Toru Ide, Minako Hirano, Daiki Yamamoto, Mami Asakura, Yuki Kitamura
    • Organizer
      The 58th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2020 Research-status Report
    • Int'l Joint Research
  • [Presentation] A gold nano-electrode for single channel detection.2019

    • Author(s)
      Toru Ide, Minako Hirano, Kota Kaneko, Huimin Ma
    • Organizer
      The 57th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2019 Research-status Report
  • [Presentation] Wide-field single-molecule fluorescence polarization detection by hybrid photo-detectors (HPDs)2019

    • Author(s)
      Atsuhito Fukasawa, Gaku Nakano, Minako Hirano, Toru Ide, Hiroaki Yokota
    • Organizer
      The 57th Annual Meeting of the Biophysical Society of Japan
    • Related Report
      2019 Research-status Report
  • [Presentation] 新規な殺蚊Cry46Abトキシンの殺虫活性メカニズム2019

    • Author(s)
      早川 徹,榊原 暁,武部 聡,井出 徹
    • Organizer
      第63回日本応用動物昆虫学会大会
    • Related Report
      2019 Research-status Report
  • [Presentation] Wide-field single-molecule multicolor fluorescence detection by hybrid photo-detectors (HPDs)2018

    • Author(s)
      Atsuhito Fukasawa, Gaku Nakano, Hiroaki Yokota, Minako Hirano, Toru Ide
    • Organizer
      日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] A simple method for single ion channel recordings.2018

    • Author(s)
      Kota Kaneko, Huimin Ma, Minako Hirano, Toru Ide
    • Organizer
      日本生物物理学会年会
    • Related Report
      2018 Research-status Report
  • [Presentation] Tomoya Ishido, Toru Ide, Minako Hirano2018

    • Author(s)
      Modifications of K+ channel property.
    • Organizer
      日本生物物理学会年会
    • Related Report
      2018 Research-status Report

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Published: 2018-04-23   Modified: 2024-12-25  

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