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Development and AFM observation of immobilized planar membranes with controlled orientation of membrane proteins and asymmetry of lipid membranes

Research Project

Project/Area Number 18K19422
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 48:Biomedical structure and function and related fields
Research InstitutionKanazawa University

Principal Investigator

Sumino Ayumi  金沢大学, ナノ生命科学研究所, 助教 (80717140)

Project Period (FY) 2018-06-29 – 2022-03-31
Project Status Completed (Fiscal Year 2021)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2020: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2019: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords高速原子間力顕微鏡 / 非対称膜 / 非対称脂質二分子膜 / イオンチャネル / 原子間力顕微鏡 / 固定化平面膜 / 膜タンパク質
Outline of Final Research Achievements

The purpose of this study is to develop a method to prepare supported lipid bilayers with simultaneous control of membrane protein orientation and lipid membrane asymmetry, and to observe the dynamics of membrane proteins whose function is related to lipid membrane asymmetry by high-speed AFM. By FY 2019, planar membranes were prepared on AFM substrates by the droplet contact method. Using a micromanipulator and a microinjector on the sample stage of a tip-scanning high-speed AFM, we were able to prepare a uniform lipid film on the AFM substrate. However, there was a problem that the optical system of the tip-scanning high-speed AFM was unstable. Therefore, in FY2021, we replaced the tip-scanning high-speed AFM with a more stable sample-scanning high-speed AFM and adjusted it to a level where it could operate stably.

Academic Significance and Societal Importance of the Research Achievements

AFM基板上での膜タンパク質の配向と脂質膜の非対称性の同時制御ができれば、膜タンパク質の機能動態に及ぼす脂質の非対称性の影響などについて分子レベルでの議論が可能になる。

Report

(5 results)
  • 2021 Annual Research Report   Final Research Report ( PDF )
  • 2020 Research-status Report
  • 2019 Research-status Report
  • 2018 Research-status Report

Research Products

(13 results)

All 2021 2020 2019

All Journal Article (7 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 7 results,  Open Access: 5 results) Presentation (6 results) (of which Invited: 2 results)

  • [Journal Article] Antibacterial Activity of Membrane-Permeabilizing Bactericidal Cyclodextrin Derivatives2021

    • Author(s)
      H. Yamamura, T. Hagiwara, Y. Hayashi, K. Ogawa, H. Kato T. Katsu K. Masuda A. Sumino H. Yamashita R. Jinno M. Abe A. Miyagawa
    • Journal Title

      ACS Omega

      Volume: 6(47) Pages: 31831-31842

    • DOI

      10.1021/acsomega.1c04541

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Macrocyclic Peptide-Conjugated Tip for Fast and Selective Molecular Recognition Imaging by High-Speed Atomic Force Microscopy2021

    • Author(s)
      L. Pupplin, D. Kanayama, N. Terasaka, K. Sakai, N. Kodera, K. Umeda, A. Sumino, M. Arin, W. Wei, H. Tanaka, T. Fukuma, H. Suga, K. Matsumoto, M. Shibata
    • Journal Title

      ACS Appl. Mater. Interfaces

      Volume: 13(46) Pages: 54817-54829

    • DOI

      10.1021/acsami.1c17708

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Visualisation of helical structures of poly(diphenylacetylene)s bearing chiral amide pendants by atomic force microscopy2021

    • Author(s)
      A. Yurtsever, S. Das, T. Nishimura, R. Rodriguez, D. Hirose, K. Miyata, A. Sumino, T. Fukuma, K. Maeda
    • Journal Title

      Chemical Communications

      Volume: 57(92) Pages: 12266-12269

    • DOI

      10.1039/d1cc05341h

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Essential Requirements of Biocompatible Cellulose Solvents2021

    • Author(s)
      T. Komori, H. Satria, K. Miyamura, A. Ito, M. Kamiya, A. Sumino, T. Onshi, K. Ninomiya, K. Takahashi, J. Anderson, T. Uto, K. Kuroda
    • Journal Title

      ACS Sustainable Chemistry & Engineering

      Volume: 9(35) Pages: 11825-11836

    • DOI

      10.1021/acssuschemeng.1c03438

    • Related Report
      2021 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Plasma membrane anchored nanosensor for quantifying endogenous production of H2O2 in living cells2021

    • Author(s)
      Hosogi Shigekuni、Marunaka Yoshinori、Ashihara Eishi、Yamada Tadaaki、Sumino Ayumi、Tanaka Hideo、Puppulin Leonardo
    • Journal Title

      Biosens Bioelectron

      Volume: 179 Pages: 113077-113077

    • DOI

      10.1016/j.bios.2021.113077

    • Related Report
      2020 Research-status Report
    • Peer Reviewed
  • [Journal Article] Urea-assisted reconstitution of discoidal high-density lipoprotein2020

    • Author(s)
      Fukuda, R.; Saito, M.; Shibukawa, S.; Sumino, A.; Nakano, M.; Murakami, T.
    • Journal Title

      Biochemistry

      Volume: 59 Pages: 1455-1464

    • DOI

      10.1021/acs.biochem.0c00075

    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [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 Pages: 1-10

    • DOI

      10.1126/sciadv.aax0495

    • NAID

      120006649788

    • Related Report
      2019 Research-status Report
    • Peer Reviewed / Open Access
  • [Presentation] Structural Dynamics of The Cytoplasmic Domain of Transient Receptor Potential Vanilloid1(TRPV1) in Lipid Bilayer Observed by High Speed Atomic Force Microscopy (HS-AFM)2021

    • Author(s)
      D. Mukai, Y. Zhao, M. Shibata, M. Hattori, A. Sumino
    • Organizer
      第44回日本分子生物学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 高速原子間力顕微鏡による膜中TRPV1 チャネルの動態観察2021

    • Author(s)
      向 大地, Yimeng Zhao, 柴田 幹大, 服部 素之, 角野 歩
    • Organizer
      第59回日本生物物理学会年会
    • Related Report
      2021 Annual Research Report
  • [Presentation] 高速AFMによるハブ毒液由来脂質分解酵素PLA2の膜認識機構の解明2020

    • Author(s)
      神谷 孫斗, 柴田 幹大, 上田 直子, 角野 歩
    • Organizer
      第58回日本生物物理学会年会
    • Related Report
      2020 Research-status Report
  • [Presentation] Studying ion channels in reconstituted membrane using atomic force microscopy2019

    • Author(s)
      角野 歩、炭竈 享司、内橋 貴之、老木 成稔
    • Organizer
      日本生物物理学会第56回年会
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] 高速AFMによるサソリ毒ペプチドとK+チャネルの結合動態解析2019

    • Author(s)
      角野 歩
    • Organizer
      大阪大学蛋白質研究所セミナー
    • Related Report
      2018 Research-status Report
    • Invited
  • [Presentation] 原子間力顕微鏡のイオンチャネル研究への応用2019

    • Author(s)
      角野 歩
    • Organizer
      第3回イオンチャネル研究会
    • Related Report
      2018 Research-status Report

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Published: 2018-07-25   Modified: 2023-01-30  

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