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two-dimensional construction of ECM-glycoconjugate chips using single molecule manipulation by glycosyltransferase attached with AFM tip

Research Project

Project/Area Number 24655098
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeSingle-year Grants
Research Field Polymer chemistry
Research InstitutionTokyo Institute of Technology

Principal Investigator

MORI Toshiaki  東京工業大学, 生命理工学研究科, 准教授 (50262308)

Project Period (FY) 2012-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords1分子計測 / 糖転移反応 / AFM / フォースカーブ測定 / 糖鎖チップ / グライコミクス / 糖転移酵素 / SPM / コンドロイチン合成酵素
Research Abstract

In order to construct ECM-glycoconjugate chips, we employed the single molecule manipulation methods . First, we examine the preparation of AFM tips modified with glycosyltransferase. As a result, we could prepare the enzyme tips. And then, we performed the force-distance measurements with the enzyme tip to the substrates on the mica surface. We examined the interaction between enzymes and the corresponding substrates under several conditions. We confirm the detection of the procedures for the enzymatic reaction. And we could control the movoment of the tip in the area with 50 nm width.

Report

(3 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Research-status Report
  • Research Products

    (20 results)

All 2014 2013 2012 2011 Other

All Journal Article (9 results) (of which Peer Reviewed: 9 results) Presentation (11 results) (of which Invited: 2 results)

  • [Journal Article] Single-Molecular Enzymatic Elongation of Hyaluronan Polymers Visualized by High-Speed Atomic Force Microscopy2013

    • Author(s)
      Toshiaki Mori, Atsushi Hirose, Tatsuya Hagiwara, Masanori Ohtsuka, Yoshimitsu Kakuta, Koji Kimata, and Yoshio Okahata
    • Journal Title

      J. Am. Chem. Soc.

      Volume: 134 Pages: 20254-20257

    • Related Report
      2013 Final Research Report 2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Kinetic monitoring of site-directed mutationalβ-amylase catalysis on a 27-MHz QCM2012

    • Author(s)
      Toshiaki Mori, Masayoshi Shibata, Takanori Nihira, Bunzo Mikami, and Yoshio Okahata
    • Journal Title

      J. Mol. Cat. B : Enzyme

      Volume: 82 Pages: 121-126

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Kinetics of Iterative Carbohydrate Transfer to Polysaccharide Catalyzed by Chondroitin Polymerase on a Highly Sensitive Flow-Type 27 MHz Quartz-Crystal Microbalance2012

    • Author(s)
      Toshiaki Mori, Takayuki Kodera, Hiroshi Yoshimine, Yoshimitsu Kakuta, Nobuo Sugiura, Koji Kimata, and Yoshio Okahata
    • Journal Title

      Chem. Eur. J.

      Volume: 18(Highlight (Cover)) Pages: 7388-7393

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Kinetics of iterative carbohydrate transfer to polysaccharide catalyzed by chondroitin polymerase on a highly sensitive flow-type 27 MHz quartz-crystal microbalance.2012

    • Author(s)
      Mori T, Kodera T, Yoshimine H, Kakuta Y, Sugiura N, Kimata K, Okahata Y.
    • Journal Title

      Chemistry

      Volume: 18 Issue: 24 Pages: 7388-7393

    • DOI

      10.1002/chem.201200342

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Kinetic monitoring of site-directed mutational β-amylase catalysis on a 27-MHz QCM2012

    • Author(s)
      Toshiaki Mori, Masayoshi Shibata, Takanori Nihira, Bunzo Mikami, and Yoshio Okahata
    • Journal Title

      J. Mol. Cat. B: Enzyme

      Volume: 82 Pages: 121-126

    • DOI

      10.1016/j.molcatb.2012.05.019

    • Related Report
      2012 Research-status Report
    • Peer Reviewed
  • [Journal Article] Single-Molecule Force Spectroscopy for Studying Kinetics of Enzymatic Dextran Elongations2011

    • Author(s)
      Toshiaki Mori, Megumi Asakura, and Yoshio Okahata
    • Journal Title

      J. Am. Chem. Soc.

      Volume: 133 Pages: 5701-5703

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Kinetic Studies of Dextransucrase Enzyme Reactions on a Substrate or Enzyme-Immobilized 27 MHz Quartz Crystal Microbalance2011

    • Author(s)
      Takanori Nihira, Toshiaki Mori, Megumi Asakura, and Yoshio Okahata
    • Journal Title

      Langmuir

      Volume: 27 Pages: 2107-2111

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Single-Molecule Force Spectroscopy for Kinetics of Enzymatic Elongation of glycoconjugate2011

    • Author(s)
      Toshiaki Mori, Megumi Asakura, and Yoshio Okahata
    • Journal Title

      Glycoconjugate J.

      Volume: 28 Pages: 292-292

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Binding Kinetics of Lectins to Sugar Surfaces on a Quartz-Crystal Microbalance

    • Author(s)
      Yoshio Okahata and Toshiaki Mori
    • Journal Title

      Methods in Molecular Biology

      Volume: (in press)

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Presentation] 高速原子間力顕微鏡を用いたグリコサミノグリカン糖鎖伸長反応の1分子観察2014

    • Author(s)
      森俊明
    • Organizer
      日本結合組織学会
    • Place of Presentation
      ウインクあいち
    • Related Report
      2013 Final Research Report
    • Invited
  • [Presentation] AFMフォースマッピングを用いた酵素的キチン分解反応の加速効果の直接観察2014

    • Author(s)
      宍戸啓介、中川裕子、森俊明
    • Organizer
      日本化学会第94春季年会
    • Place of Presentation
      名古屋大学
    • Related Report
      2013 Final Research Report
  • [Presentation] AFMを用いた脂質二分子膜中の膜タンパク質の動的挙動解析2014

    • Author(s)
      田中利奈、森俊明
    • Organizer
      日本化学会第94春季年会
    • Place of Presentation
      名古屋大学
    • Related Report
      2013 Final Research Report
  • [Presentation] 高速AFMを用いた糖脂質膜上での糖転移酵素反応の直接イメージング2014

    • Author(s)
      小林真也、森俊明
    • Organizer
      日本化学会第94春季年会
    • Place of Presentation
      名古屋大学
    • Related Report
      2013 Final Research Report
  • [Presentation] 原子間力顕微鏡を用いたグリコサミノグリカン糖鎖伸長反応の一分子直接観察2014

    • Author(s)
      森 俊明
    • Organizer
      第46回日本結合組織学会学術大会・第61回マトリックス研究会大会 合同学術集会
    • Place of Presentation
      ウインクあいち
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] 生体膜上のナノ糖鎖の生成に関する精密計測2013

    • Author(s)
      森俊明
    • Organizer
      高分子学会九州若手の会
    • Place of Presentation
      アルモニーサンク小倉
    • Year and Date
      2013-07-05
    • Related Report
      2013 Final Research Report
  • [Presentation] ベロ毒素修飾チップによる糖脂質ラフト界面の分子マッピング2013

    • Author(s)
      森俊明、小泉翔平、露木由実
    • Organizer
      第62回高分子討論会
    • Place of Presentation
      金沢大学
    • Related Report
      2013 Final Research Report
  • [Presentation] ベロ毒素修飾チップによるGb3Cer含有脂質膜のフォースマッピング観察2013

    • Author(s)
      森俊明、小泉翔平、岡畑恵雄
    • Organizer
      バイオ高分子シンポジウム2013
    • Place of Presentation
      東工大
    • Related Report
      2013 Final Research Report
  • [Presentation] ベロ毒素修飾探針を用いたAFMフォースマッピングによる脂質膜中におけるGb3セラミドの分布観察2013

    • Author(s)
      森俊明、小泉翔平、岡畑恵雄
    • Organizer
      第62回高分子学会年次大会
    • Place of Presentation
      京都国際会議場
    • Related Report
      2013 Final Research Report
  • [Presentation] ベロ毒素修飾探針を用いたAFMフォースマッピングによる脂質膜中におけるGb3セラミドの分布観察2013

    • Author(s)
      森 俊明、小泉翔平、岡畑 恵雄
    • Organizer
      第62回高分子学会年次大会
    • Place of Presentation
      京都国際会議場
    • Related Report
      2013 Annual Research Report
  • [Presentation] ベロ毒素修飾チップによる糖脂質ラフト界面の 分子マッピング2013

    • Author(s)
      森 俊明、小泉翔平、露木由実
    • Organizer
      第62回高分子討論会
    • Place of Presentation
      金沢大学
    • Related Report
      2013 Annual Research Report

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Published: 2013-05-31   Modified: 2019-07-29  

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