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Reconstitution physiology: Functional construction of channel and membrane

Research Project

Project/Area Number 17H04017
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionUniversity of Fukui

Principal Investigator

Oiki Shigetoshi  福井大学, 高エネルギー医学研究センター, 特命教授 (10185176)

Co-Investigator(Kenkyū-buntansha) 岩本 真幸  福井大学, 学術研究院医学系部門, 教授 (40452122)
Project Period (FY) 2017-04-01 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000)
Fiscal Year 2019: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2018: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2017: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
Keywords脂質2重膜 / 接触バブル2重膜法 / 膜張力 / イオンチャネル / コレステロール / 脂質二重膜 / 一分子測定 / 膜物性 / in vitro転写翻訳 / ゲーティング / 機械受容性チャネル / インビトロ転写ー発現系 / 脂質平面膜 / 膜再構成 / 電気生理学 / 膜物理的特性
Outline of Final Research Achievements

Cell membrane is a core platform for the cellular activity, in which ion channel proteins, embedded in the membrane, operate their function through interaction with the membrane. However, cell membrane has complex structure, and elucidating molecular mechanism of channel-membrane interaction is very difficult. To circumvent this difficulty, we established a simple lipid bilayer membrane, in which channel proteins were incorporated. We developed a method to synthesize channel protein in this lipid bilayer system, where newly synthesized channel proteins were transferred into the membrane and exhibited channel activities. Changes in chemical composition of the membrane and force generated in the membrane were reproduced, and we found that channel are subject to change its activity by membrane stretch under quantitative evaluation of the membrane tension. Effect of various types of native lipids and other hydrophobic substances were examined for the molecular mechanism.

Academic Significance and Societal Importance of the Research Achievements

生理学は「細胞や組織をありのままで観察することでその機能を明らかにする」という伝統的手法が使われてきたが、複雑系である生体の機能を明らかにするにはこのような方法だけでは不十分である。このようなトップダウンとは逆の「ボトムアップアプローチを行うことによって生体機能を明らかにする」というコンセプトに基づくのが本研究の学術的意義である。そしてこのようなアプローチを行うには方法論が不可欠であるが、そのための基礎的方法を独自に開発したことが本研究の戦略となる。この方法をもとに、生体膜の複雑性を次々に再現し、この実験系ならではの解析力によってチャネルと膜の相互作用についてその分子機構を明らかにした。

Report

(4 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Annual Research Report
  • 2017 Annual Research Report
  • Research Products

    (27 results)

All 2020 2019 2018 2017

All Journal Article (17 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 17 results,  Open Access: 8 results,  Acknowledgement Compliant: 2 results) Presentation (5 results) (of which Int'l Joint Research: 1 results,  Invited: 5 results) Book (1 results) Patent(Industrial Property Rights) (4 results)

  • [Journal Article] 脂質2重膜の水透過2020

    • Author(s)
      老木成稔
    • Journal Title

      腎と透析

      Volume: 88 Pages: 322-326

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Queueing arrival and release mechanism for K+ permeation through a potassium channel.2019

    • Author(s)
      Sumikama, T. and Oiki, S.
    • Journal Title

      J. Physiol. Sci.

      Volume: 69 Issue: 6 Pages: 919-930

    • DOI

      10.1007/s12576-019-00706-4

    • NAID

      120006715810

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] High-speed AFM reveals accelerated binding of agitoxin-2 to a K+ channel by induced fit2019

    • Author(s)
      A. Sumino,T. Sumikama,T. Uchihashi,S. Oiki
    • Journal Title

      Science Advances

      Volume: 5 Issue: 7 Pages: 1-10

    • DOI

      10.1126/sciadv.aax0495

    • NAID

      120006649788

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Drop-in-well chamber for droplet interface bilayer with built-in electrodes2019

    • Author(s)
      Urakubo Kazuhiro、Iwamoto Masayuki、Oiki Shigetoshi
    • Journal Title

      Methods Enzymol.

      Volume: 621 Pages: 347-363

    • DOI

      10.1016/bs.mie.2019.02.012

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] In bulla functional channel expression systems that mimic bacterial synthetic membranes2019

    • Author(s)
      Iwamoto Masayuki、Oiki Shigetoshi
    • Journal Title

      Methods Enzymol.

      Volume: 621 Pages: 231-244

    • DOI

      10.1016/bs.mie.2019.02.011

    • ISBN
      9780128181171
    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A concise method for quantitative analysis of interactions between lipids and membrane proteins2019

    • Author(s)
      Inada Masataka、Kinoshita Masanao、Sumino Ayumi、Oiki Shigetoshi、Matsumori Nobuaki
    • Journal Title

      Analytica Chimica Acta

      Volume: 1059 Pages: 103-112

    • DOI

      10.1016/j.aca.2019.01.042

    • Related Report
      2019 Annual Research Report 2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Lipid Bilayer Experiments with Contact Bubble Bilayers for Patch-Clampers2019

    • Author(s)
      Iwamoto Masayuki、Oiki Shigetoshi
    • Journal Title

      J. Vis. Exp.

      Volume: 143 Issue: 143

    • DOI

      10.3791/58840

    • Related Report
      2019 Annual Research Report 2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Constitutive boost of a K+ channel via inherent bilayer tension and a unique tension-dependent modality2018

    • Author(s)
      Iwamoto Masayuki、Oiki Shigetoshi
    • Journal Title

      Proc. Natl. Acad. Sci. U. S. A.

      Volume: 115 Issue: 51 Pages: 13117-13122

    • DOI

      10.1073/pnas.1812282115

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Thylakoid membranes contain a non-selective channel permeable to small organic molecules2018

    • Author(s)
      Kojima S, Iwamoto M, Oiki S, Tochigi S, Takahashi H
    • Journal Title

      Journal of Biological Chemistry

      Volume: 印刷中 Issue: 20 Pages: 7777-7785

    • DOI

      10.1074/jbc.ra118.002367

    • Related Report
      2018 Annual Research Report 2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Concurrent In Vitro Synthesis and Functional Detection of Nascent Activity of the KcsA Channel under a Membrane Potential2018

    • 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

    • Related Report
      2018 Annual Research Report 2017 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Structure and dynamics of solvent molecules inside the polytheonamide B channel in different environments: a molecular dynamics study2018

    • Author(s)
      Kalathingal Mahroof、Sumikama Takashi、Mori Toshifumi、Oiki Shigetoshi、Saito Shinji
    • Journal Title

      Physical Chemistry Chemical Physics

      Volume: 20 Issue: 5 Pages: 3334-3348

    • DOI

      10.1039/c7cp06299k

    • Related Report
      2018 Annual Research Report 2017 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Lipid Bilayers Manipulated through Monolayer Technologies for Studies of Channel-Membrane Interplay2018

    • Author(s)
      Oiki S, Iwamoto M
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 41 Issue: 3 Pages: 303-311

    • DOI

      10.1248/bpb.b17-00708

    • NAID

      130006407489

    • ISSN
      0918-6158, 1347-5215
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Entrance of Channel Determines Ion Permeation Rate through the Kv1.2 Channel2018

    • Author(s)
      炭竈 享司、老木 成稔
    • Journal Title

      Seibutsu Butsuri

      Volume: 58 Issue: 1 Pages: 012-016

    • DOI

      10.2142/biophys.58.012

    • NAID

      130006322043

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Channel Formation and Membrane Deformation via Sterol-Aided Polymorphism of Amphidinol 32017

    • 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

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Membrane Perfusion of Hydrophobic Substances Around Channels Embedded in the Contact Bubble Bilayer2017

    • Author(s)
      Iwamoto M, Oiki S
    • Journal Title

      Scientific Reports

      Volume: 7 Issue: 1 Pages: 6857-6857

    • DOI

      10.1038/s41598-017-07048-4

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Oriented Reconstitution of the Full-Length KcsA Potassium Channel in a Lipid Bilayer for AFM Imaging2017

    • Author(s)
      Ayumi Sumino, Takayuki Uchihashi and Shigetoshi Oiki
    • Journal Title

      J. Phys. Chem. Lett.

      Volume: 8 Issue: 4 Pages: 785-793

    • DOI

      10.1021/acs.jpclett.6b03058

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] 生物物理学会における多様性2017

    • Author(s)
      豊島 有、飯野 雄一
    • Journal Title

      Seibutsu Butsuri

      Volume: 57 Issue: 1 Pages: 001-001

    • DOI

      10.2142/biophys.57.001

    • NAID

      130005296440

    • ISSN
      0582-4052, 1347-4219
    • Related Report
      2017 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Presentation] イオンチャネル一分子研究の再構成膜系からのアプローチ2020

    • Author(s)
      老木成稔
    • Organizer
      生理研研究会
    • Related Report
      2019 Annual Research Report
    • Invited
  • [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(Effects of cholesterol on the function of the KcsA potassium channel)2017

    • Author(s)
      岩本真幸、老木成稔
    • Organizer
      第94回日本生理学会大会(アクトシティ浜松、浜松)
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] 脂質二重膜がカリウムチャネル開閉に及ぼす多様な影響(Specific and non-specific actions of membrane lipids on the gating of the potassium channel)2017

    • Author(s)
      岩本真幸、老木成稔
    • Organizer
      日本生物物理学会第55回年会(熊本大学、熊本)シンポジウム「いろんなスケールで働く膜タンパク質の作動原理:実験と理論の新展開(Operating principles of membrane proteins at multiscale resolutions)」
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] KcsAカリウムチャネル膜相と作用のための一分子実験法2017

    • Author(s)
      老木成稔
    • Organizer
      蛋白研セミナー
    • Related Report
      2017 Annual Research Report
    • Invited
  • [Presentation] Ion Permeation and Gating of the KcsA Potassium Channel Reconstituted in the Manipulated Membrane.2017

    • Author(s)
      Oiki, S.
    • Organizer
      Ion Channel Modulation Symposium, Cambridge, UK
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research / Invited
  • [Book] Reference Module in Life Sciences2017

    • Author(s)
      Shigetoshi Oiki
    • Total Pages
      43
    • Publisher
      Elsevier
    • ISBN
      9780128096338
    • Related Report
      2017 Annual Research Report
  • [Patent(Industrial Property Rights)] 脂質二重膜の水透過性の評価システム、脂質二重膜の水透過性の評価方法、および、脂質二重膜の水透過性を制御する薬剤のスクリーニング方法2019

    • Inventor(s)
      老木 成稔・岩本真幸・矢野啓太
    • Industrial Property Rights Holder
      老木 成稔・岩本真幸・矢野啓太
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2019-012092
    • Filing Date
      2019
    • Related Report
      2018 Annual Research Report
  • [Patent(Industrial Property Rights)] 接触液胞2重膜法による膜物性変化の実時間測定法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
      特許
    • Industrial Property Number
      2017-246976
    • Filing Date
      2017
    • Related Report
      2017 Annual Research Report
  • [Patent(Industrial Property Rights)] 脂質二重膜形成装置、脂質二重膜形成方法、および評価システム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
      特許
    • Industrial Property Number
      2017-063796
    • Filing Date
      2017
    • Related Report
      2017 Annual Research Report
  • [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
    • Related Report
      2017 Annual Research Report

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Published: 2017-04-28   Modified: 2021-02-19  

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