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Anlyses of enzymatic reaction about glycoconjugates using Atomic Force Microscopy in single molecule level

Research Project

Project/Area Number 23350078
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Chemistry related to living body
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)

Co-Investigator(Kenkyū-buntansha) 高橋 俊太郎  甲南大学, 付置研究所, 講師 (40456257)
古澤 宏幸  東京工業大学, 生命理工学研究科, 助教 (60345395)
川崎 剛美  東京工業大学, 生命理工学研究科, 助教 (60334504)
Project Period (FY) 2011-04-01 – 2014-03-31
Project Status Completed (Fiscal Year 2013)
Budget Amount *help
¥19,240,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥4,440,000)
Fiscal Year 2013: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2012: ¥5,590,000 (Direct Cost: ¥4,300,000、Indirect Cost: ¥1,290,000)
Fiscal Year 2011: ¥8,060,000 (Direct Cost: ¥6,200,000、Indirect Cost: ¥1,860,000)
Keywords原子間力顕微鏡 / 糖鎖 / 細胞膜 / 1分子計測 / 糖鎖関連酵素 / キチン / キチン分解酵素 / 力学計測 / 分子マッピング / 糖鎖伸長反応 / 糖転移酵素 / 一分子計測 / 高速AFM / 力学計測法 / 高感度水晶発振子マイクロバランス
Research Abstract

Atomic force microscopy (AFM) can detect forces as intermolecular interaction forces by force-curve measurements. For example, when a ligand is immobilized on a chip, and a receptor is immobilized on a plate, we can measure single-molecular interaction forces between two molecules. The distribution of glycolipid microdomain in biomembrane is difficult to observe in nanometer scale. We measured a distribution of glycolipid Gb3Ceramide (Gb3Cer) in lipid bilayer by AFM force mapping measurement using Verotoxin modified tip.
In this study, we have analyzed interaction forces between a verotoxin that is a substance responsible for o-157 and Gb3 sugar molecules on a cell surface. The rupture forces were obtained by changing loading rates and we can get an effective bond length which is an important parameter for an analysis of interaction mechanisms. We detected single-molecular interaction forces, and we discussed about the interaction mechanism in more detail by an analysis of the forces.

Report

(4 results)
  • 2013 Annual Research Report   Final Research Report ( PDF )
  • 2012 Annual Research Report
  • 2011 Annual Research Report
  • Research Products

    (29 results)

All 2014 2013 2012 2011 Other

All Journal Article (9 results) (of which Peer Reviewed: 9 results) Presentation (20 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
    • 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

    • 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 Pages: 7388-7393

    • Related Report
      2013 Final Research Report
    • Peer Reviewed
  • [Journal Article] Single-Molecular Enzymatic Elongation of Hyaluronan Polymers Visualized by High-Speed Atomic Force Microscopy2012

    • 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 Issue: 50 Pages: 20254-20257

    • DOI

      10.1021/ja309646s

    • Related Report
      2012 Annual Research 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 2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Single-Molecule Force Spectroscopy for Kinetics of Enzymatic Elongation of glycoconjutate2011

    • 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] 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 Issue: 15 Pages: 5701-5703

    • DOI

      10.1021/ja200094f

    • Related Report
      2011 Annual 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] 種々のキチン分解酵素によるキチンへの結合・分解挙動のAFM1分子解析2014

    • Author(s)
      加藤早紀, 中川裕子, 森俊明
    • Organizer
      第63回高分子討論会
    • Place of Presentation
      長崎大学
    • Related Report
      2013 Final Research Report
  • [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] キチナーゼ修飾探針を用いたキチンフィルムの酵素的ナノパターニング法の開発2014

    • Author(s)
      宍戸啓介, 中川裕子, 森俊明
    • Organizer
      第63回高分子学会年次大会
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2013 Final Research Report
  • [Presentation] キチン分解酵素ChiA, ChiBおよびChiCのキチンへの結合解離メカニズムの1分子解析2014

    • Author(s)
      加藤早紀, 中川裕子, 森俊明
    • Organizer
      第63回高分子学会年次大会
    • Place of Presentation
      名古屋国際会議場
    • Related Report
      2013 Final Research Report
  • [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] キチン分解酵素ChiA, ChiBおよびChiCのキチンへの結合・分解挙動の1分子観察2014

    • Author(s)
      加藤早紀, 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] 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] キチン分解酵素ChiA,ChiB およびChiC の キチンへの結合解離挙動の1 分子観察2014

    • Author(s)
      加藤早紀・中川裕子・森 俊明
    • Organizer
      第94日本化学会春季年会
    • Place of Presentation
      名古屋大学
    • Related Report
      2013 Annual Research Report
  • [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] キチナーゼ修飾探針を用いたAFMフォースマッピングによるキチンフィルムの結合・分解挙動の観察2013

    • Author(s)
      宍戸啓介, 中川裕子, 森俊明
    • Organizer
      第62回高分子討論会
    • 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
    • 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 Annual Research Report 2013 Final Research Report
  • [Presentation] キチナーゼ修飾探針を用いたAFMフォースマッピングによるキチンフィルムへの結合・解離挙動の直接観察2013

    • Author(s)
      宍戸啓介, 森俊明, 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] キチナーゼ修飾探針を用いたAFMフォースマッピングによる キチンフィルムへの結合・分解挙動の直接観察2013

    • Author(s)
      宍戸 啓介・森 俊明・岡畑 惠雄
    • 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Gb3セラミド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 惠雄
    • Organizer
      バイオ高分子シンポジウム2013
    • Place of Presentation
      東工大大岡山キャンパス
    • Related Report
      2013 Annual Research Report
  • [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
      小倉
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] キチナーゼ修飾探針を用いたAFMフォースマッピングによる キチンフィルムへの結合・分解挙動の観察2013

    • Author(s)
      宍戸啓介・中川裕子・森 俊明
    • Organizer
      第62回高分子討論会
    • Place of Presentation
      金沢大学
    • 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: 2011-04-06   Modified: 2019-07-29  

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