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Development of Non-Supported Membrane over Nanowells for Visualization of Functional Dynamics of Membrane Proteins with High-Speed AFM

Research Project

Project/Area Number 26600052
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Nanobioscience
Research InstitutionKanazawa University

Principal Investigator

UCHIHASHI TAKAYUKI  金沢大学, 数物科学系, 教授 (30326300)

Co-Investigator(Kenkyū-buntansha) IINO Ryota  自然科学研究機構, 岡崎統合バイオサイエンスセンター, 教授 (70403003)
Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords一分子計測 / 膜タンパク質 / 高速原子間力顕微鏡 / 1分子イメージング・ナノ計測
Outline of Final Research Achievements

We prepared nano-pore arrays with a diameter of 100nm on glass and silicon substrate using electron-beam lithography and focused ion beam. Also we succeeded in sealing the nano pores with lipid bilayers. Purple membranes in which two dimensional crystals of bacteriorhodopsin are embedded were used as a test sample and the individual bacteriorhodopsin molecules were resolved on the nano pore with high-speed AFM. We also developed a method to conveniently prepare the nano pores on a silicone elastomer using small beads with a diameter of 100nm as a template. We observed ion-channel membrane protein KcsA on a supported lipid bilayer and visualized individual KcsA tetramers and theirs diffusion dynamics in the membrane. As the next step, we will reconstitute KcsA molecules into the membrane on the nano pore and reveal structural dynamics of KcsA..

Report

(3 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • Research Products

    (22 results)

All 2016 2015 2014

All Journal Article (8 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Acknowledgement Compliant: 2 results) Presentation (12 results) (of which Int'l Joint Research: 3 results,  Invited: 12 results) Book (2 results)

  • [Journal Article] Functional extension of high-speed AFM for wider biological applications.2016

    • Author(s)
      Uchihashi T, Watanabe H, Fukuda S, Shibata M, Ando T.
    • Journal Title

      Ultramicroscopy

      Volume: 160 Pages: 182-196

    • DOI

      10.1016/j.ultramic.2015.10.017

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Potential pre-pore trimer formation by the Bacillus thuringiensis mosquito-specific toxin: Molecular insights into a critical prerequisite of membrane-bound monomers2015

    • Author(s)
      Wilaiwan Sriwimol, Aratee Aroonkesorn, Somsri Sakdee, Chalermpol Kanchanawarin, Takayuki Uchihashi, Toshio Ando and Chanan Angsuthanasombat
    • Journal Title

      The Journal of Biological Chemistry

      Volume: 290 Issue: 34 Pages: 20793-20803

    • DOI

      10.1074/jbc.m114.627554

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Method of mechanical holding of cantilever chip for tip-scan high-speed atomic force microscope.2015

    • Author(s)
      Fukuda S, Uchihashi T, Ando T.
    • Journal Title

      Rev Sci Instrum.

      Volume: 86 Issue: 6 Pages: 063703-063703

    • DOI

      10.1063/1.4922381

    • NAID

      120005647653

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Long-tip high-speed atomic force microscopy for nanometer-scale imaging in live cells2015

    • Author(s)
      M. Shibata, T. Uchihashi, T. Ando and R. Yasuda
    • Journal Title

      Scientific Reports

      Volume: 5 Pages: 8724-8724

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] Probing structural dynamics of an artificial protein cage using high-speedatomic force microscopy2015

    • Author(s)
      M. Imamura, T. Uchihashi, T. Ando, A. Leifert, U. Simon, A. D. Malay and J. G. Heddle
    • Journal Title

      Nano Letters

      Volume: 15 Issue: 2 Pages: 1331-1335

    • DOI

      10.1021/nl5045617

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] Two-way traffic of glycoside hydrolase family 18 processive chitinases on crystalline chitin2014

    • Author(s)
      K. Igarashi, T. Uchihashi, T. Uchiyama, H. Sugimoto, M. Wada, K. Suzuki, S. Sakuda, T. Ando, T. Watanabe, and M. Samejima
    • Journal Title

      Nature Communications

      Volume: 5 Pages: 2975-2975

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] キチナーゼによる結晶性キチンのプロセッシブ(連続的)な分解機構の解明2014

    • Author(s)
      杉本華幸, 五十嵐圭日子, 内橋貴之, 鈴木一史, 渡邉剛志
    • Journal Title

      日本応用糖質化学会誌

      Volume: 4 Pages: 107-112

    • Related Report
      2014 Research-status Report
  • [Journal Article] 高速原子間力顕微鏡による生体試料のダイナミクス観察2014

    • Author(s)
      内橋貴之
    • Journal Title

      膜誌

      Volume: 39 Pages: 322-328

    • NAID

      130005069445

    • Related Report
      2014 Research-status Report
  • [Presentation] 高速原子間力顕微鏡で探る生体分子の機能ダイナミクス2016

    • Author(s)
      内橋貴之
    • Organizer
      表面科学技術研究会2016 生きている固体と界面 ~走査型プローブ顕微鏡で探る~
    • Place of Presentation
      神戸大学
    • Year and Date
      2016-01-21
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 高速原子間力顕微鏡で可視化する生体分子の構造ダイナミクス2015

    • Author(s)
      内橋貴之
    • Organizer
      2015年度日本物理学会北陸支部定例学術講演会
    • Place of Presentation
      金沢大学
    • Year and Date
      2015-11-28
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 高速原子間力顕微鏡で可視化するタンパク質の機能動態2015

    • Author(s)
      内橋貴之
    • Organizer
      第8回北陸合同バイオシンポジウム
    • Place of Presentation
      山中温泉、石川
    • Year and Date
      2015-10-30
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] Direct Visualization of Single Molecule Dynamics at Work with High-Speed Atomic Force Microscopy2015

    • Author(s)
      Takayuki Uchihashi
    • Organizer
      The 15th KIAS Conference on Protein Structure and Function
    • Place of Presentation
      ソウル、韓国
    • Year and Date
      2015-09-17
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Single-molecule manipulation and control of protein conformational dynamics using high-speed AFM2015

    • Author(s)
      Takayuki Uchihashi
    • Organizer
      KAKENHI International Symposium on “Studying the Function of Soft Molecular Systems”
    • Place of Presentation
      未来科学館、台場
    • Year and Date
      2015-07-09
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] High-speed atomic force microscopy: A new tool for studying protein dynamics at work2015

    • Author(s)
      Takayuki Uchihashi
    • Organizer
      IMS Asian International Symposium “Supramolecular Dynamics at the Interface of Chemistry and Biology”
    • Place of Presentation
      岡崎統合バイオ
    • Year and Date
      2015-06-12
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] 高速原子間力顕微鏡で可視化する分子動態と細胞運動2015

    • Author(s)
      内橋貴之
    • Organizer
      日本顕微鏡学会第71回学術講演会
    • Place of Presentation
      京都国際会議場
    • Year and Date
      2015-05-15
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] High-speed atomic force microscope for imaging of biomolecular dynamics at solid surface2015

    • Author(s)
      Takayuki Uchihashi
    • Organizer
      10th Annual International Electromaterials Science Symposium
    • Place of Presentation
      Wollongong, Australia
    • Year and Date
      2015-02-11
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] Visualization of single molecule dynamics at work with high-speed atomic force microscopy2015

    • Author(s)
      Takayuki Uchihashi
    • Organizer
      The 7th Biennial Australian Colloid & Interface Symposium
    • Place of Presentation
      Hobart, Tasmania, Australia
    • Year and Date
      2015-02-03
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] 高速原子間力顕微鏡によるタンパク質のダイナミクスと物性計測2015

    • Author(s)
      内橋貴之
    • Organizer
      NIBBバイオイメージングフォーラム「物理特性のイメージング」
    • Place of Presentation
      基礎生物学研究所, 愛知県
    • Year and Date
      2015-01-27
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] 高速原子間力顕微鏡の開発とバイオ応用2015

    • Author(s)
      内橋貴之
    • Organizer
      新世代研究所水和ナノ構造・界面ナノ科学合同研究会「固液界面の水和ナノ構造と生体高分子ダイナミクス」
    • Place of Presentation
      伊豆長岡温泉 天坊, 静岡県
    • Year and Date
      2015-01-25
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] High-speed atomic force microscope for studying dynamic interactions in biomolecular system2015

    • Author(s)
      Takayuki Uchihashi
    • Organizer
      The 3rd International Symposium on Dynamical Ordering of Biomolecular Systems for Creation of Integrated Functions
    • Place of Presentation
      合歓の郷,三重県
    • Year and Date
      2015-01-10
    • Related Report
      2014 Research-status Report
    • Invited
  • [Book] “光と生命の辞典” 第5章 「光による生命現象の計測」177節 高速原子間力顕微鏡2016

    • Author(s)
      内橋貴之
    • Total Pages
      486
    • Publisher
      朝倉書店
    • Related Report
      2015 Annual Research Report
  • [Book] “Development of high-speed AFM and its biological applications”, Chapter 8 in Atomic Force Microscopy in Nanobiology2014

    • Author(s)
      T. Uchihahsi, N. Kodera and T. Ando
    • Total Pages
      34
    • Publisher
      Pan Stanford Publishing
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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