生体情報伝達連鎖機構の単分子力学解析と計算機モデリング

2011 Fiscal Year Annual Research Report

• Project/Area Number: 19GS0418
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: 猪飼 篤 東京工業大学, イノベーション研究推進体, 特任教授 (50011713)
• Co-Investigator(Kenkyū-buntansha): 岸本 喜久雄 東京工業大学, 大学院・理工学研究科, 教授 (30111652) ; 長田 俊哉 東京工業大学, 大学院・生命理工学研究科, 准教授 (00201997)
• Keywords: 生体力学情報伝達 / 細胞骨格 / 生体ナノ力学 / 原子間力顕微鏡 / リンカータンパク質 / パキシリン / 有限要素法 / 分子動力学

Research Abstract

1.アクチン細胞骨格系の外部力学刺激に対する応答のレオロジー解析をおこない、細胞膜に結合した小球を引き上げる際に時間的に変化する細胞応答を解析し、無生物的な緩和現象と、生物的な応答を区別した。小球を完全に引き上げることにより、付着する細胞内タンパク質の解析を可能とした。
2.ファイバー損傷情報とこれに呼応して動員される修復タンパク質群の動向を知る実験を行った。原子間力顕微鏡を用いて、印加力を制御しつつストレスファイバーに機械的な損傷を部分的に与え、その修復可能性を検討し、修復される場合は従来観察されていたジキシンより早くパキシリンというタンパク質が傷口に集積して損傷治癒に当たることを見出した。同時に接着斑のパキソリン量が減少した。損傷が繊維を切断する大きさになると治癒は難しい。その他数種類の接着斑関連タンパク質についても検討したが、傷口への有意の集積は見られなかった。
3.アクチンストレスファイバーに直接横方向に力を印加することにより、ストレスファイバーと細胞膜結合アクトミオシン層の間につよい結合があることが力学的に証明できた。この問題は一昨年度から引き続く問題であったが、原子間力顕微鏡の弱点をいくつかの方法で補うことにより、100ナノニュートン以上の大きな力によるストレスファイバーとアクトミオシン層の動きを蛍光顕微鏡を通して確認した。任意の細胞操作を可能とする形状を持つ探針を各種開発した。
4.細胞骨格ネットワークを形成する異種タンパク質間の相互作用力を単一分子レベルのフォーススペクトルスコピー法で解析した。
5.有限要素法の細胞モデル弾性解析を行い,ストレスファイバーと細胞膜の弾性係数を逆解析し、ストレスファイバーの変形に細胞膜の変形抵抗が寄与していることを明らかにした.
6.受容体とリガンドの相互作用解析を行なうためのシステムの開発を行なった。

Research Products

• [Journal Article] Forced extension of delipidated red blood cell cytoskeleton with little indication of spectrin unfolding (2012)
  • Author(s): Afrin R, Nakaji M, Sekiguchi H, Lee D, Kishimoto K, Ikai A
  • Journal Title: Cytoskeleton Volume: 69 Pages: 101-112
  • DOI: doi:10.1002/cm.21001
  • Peer Reviewed
• [Journal Article] Interaction between pheromone and its receptor of the fission yeast Schizosaccharomyces pombe examined by a force spectroscopy stud (2012)
  • Author(s): S.Sasuga, R.Abe, O.Nikaido, S.Kiyosaki, H.Sekiguchi, A.Ikai, T.Osada
  • Journal Title: Journal of Biomedicine and Biotechnology Pages: 804793
  • DOI: doi:10.1002/cm.20537
  • Peer Reviewed
• [Journal Article] Interaction of Serum Proteins with Surface of Hemodialysis Fiber Membranes (2012)
  • Author(s): Rehana Afrin, Yuji Shirako, Kikuo Kishimoto, Atsushi Ikai
  • Journal Title: Japanese Journal of Applied Physics Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] Fabricated cantilever for AFM measurements and manipulations : Pre-stress analysis of stress fibers (2012)
  • Author(s): Shinich Machida, Takahiro Watanabe-Nakayama, Masakazu Saito, Rehana Afrin, Atsushi Ikai
  • Journal Title: Micron Volume: (印刷中)
  • Peer Reviewed
• [Journal Article] Nonlinear displacement of ventral stress fibers under externally applied lateral force by an atomic force microscope (2011)
  • Author(s): Hakari T, Sekiguchi H, Osada T, Kishimoto K, Afrin R, Ikai A
  • Journal Title: Cytoskeleton Volume: 68巻 Pages: 628-638
  • DOI: doi:10.1002/cm.20537
  • Peer Reviewed
• [Presentation] Paxillin accumulation to a damaged site on a stress fiber (2012)
  • Author(s): Takahiro Watanabe-Nakayama, Masakazu Saito, Shin-ichi Machida, Rehana Afrin, Atsushi Ikai
  • Organizer: Biophysical Society 56^<th> Annual Meeting
  • Place of Presentation: San Diego, CA, USA
  • Year and Date: 20120225-20120229
• [Presentation] Exploring Atomic Force Microscopy for Nano-medical Applications (2012)
  • Author(s): Rehana Afrin, Takahiro Watanabe-Nakayama, Shinichi Machida, Masakazu, Saito and Atsushi Ikai
  • Organizer: ACMM22/ICONN2012/APMC10 Combined Conference
  • Place of Presentation: Perth, Australia
  • Year and Date: 20120205-09
• [Presentation] AFM for Cytoskeletal Mechanics (2012)
  • Author(s): Atsushi Ikai, Rehana Afrin, Shinichi Machida, Takahiro Watanabe-Nakayama, Masakazu, Saito, Kikuo Kishimoto
  • Organizer: Linz Winter Workshop 2012
  • Place of Presentation: Lintz, Austria(invited)
  • Year and Date: 20120204-20120207
• [Presentation] Single molecule dynamic interactions between spectrin and ankyrin studied by atomic force microscopy (2012)
  • Author(s): Masakazu Saito, Takahiro W.Nakayama, Shinichi Machida, Rehana Afrin, Atsushi Ikai
  • Organizer: XIVth Annual Linz Winter Workshop
  • Place of Presentation: Linz, Austria
  • Year and Date: 20120203-20120207
• [Presentation] Biomechanics of Cytoskeletal Structures (2011)
  • Author(s): Atsushi Ikai, Rehana Afrin, Shinichi Machida, Takahiro Watanabe-Nakayama, Masakazu, Saito, Kikuo Kishimoto
  • Organizer: the Eleventh AEARU Workshop on Molecular Biology and Biotechnology
  • Place of Presentation: Hong Kong, China(invited)
  • Year and Date: 20111221-20111222
• [Presentation] Interaction of Serum Proteins with the Surface of Hemodialysis Tubing (2011)
  • Author(s): Rehana Afrin, Takahiro Watanabe-Nakayama, Shinichi Machida, Masakazu, Saito, Atsushi Ikai
  • Organizer: International Colloquium of Scanning Probe Microscopy (ICSPM19)
  • Place of Presentation: Toyako, Japan
  • Year and Date: 20111219-20111222
• [Presentation] Paxillin accumulation at strained stress fibers (2011)
  • Author(s): Takahiro Watanabe-Nakayama, Masakazu Saito, Shin-ichi Machida, Rehana Afrin, Atsushi Ikai
  • Organizer: 日本分子生物学会第34回年会
  • Place of Presentation: Yokohama, Japan
  • Year and Date: 20111213-20111216
• [Presentation] Single molecular dynamic interactions between spectrin and ankyrin studied by atomic force microscopy (2011)
  • Author(s): 齋藤勝和、中山隆宏、町田真一、レハナアフリン、猪飼篤
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川)
  • Year and Date: 20111213-20111216
• [Presentation] Paxillin accumulation at strained stress fibers (2011)
  • Author(s): 中山(渡部)隆宏、齋藤勝和、町田真一、Rehana Afrin、猪飼篤
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川)
  • Year and Date: 20111213-20111216
• [Presentation] 原子間力顕微鏡を用いたアクチンフィラメントとビンキュリンテイルの相互作用力測定 (2011)
  • Author(s): 笠原慧、中山隆宏、町田真一、斉藤勝和、長田俊哉、レハナアフリン、猪飼篤
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川)
  • Year and Date: 20111213-20111216
• [Presentation] Lateral Displacement of Stress Fibers under Direct Mechanical Manipulation (2011)
  • Author(s): Atsushi Ikai, Rehana Afrin, Shinichi Machida, Masakazu Saito, Takahiro Nakayama-Watanabe
  • Organizer: International Conference on Surface Sciences (ISSS) 6
  • Place of Presentation: Funabori, Japan
  • Year and Date: 20111211-20111215
• [Presentation] Exploring Atomic Force Microscopy for Single Cell Manipulations (2011)
  • Author(s): Rehana Afrin, Takahiro Watanabe-Nakayama, Shinichi Machida, Masakazu, Saito, Atsushi Ikai
  • Organizer: Annual Meeting of the American Society for Cell Biology
  • Place of Presentation: Denver, CO, USA
  • Year and Date: 20111203-20111207
• [Presentation] フェロモン受容体の分裂酵母細胞膜上での発現効率の研究 (2011)
  • Author(s): 金高友貴恵, 猪飼篤, 長田俊哉
  • Organizer: 第84回日本生化学会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 20110921-20110924
• [Presentation] 分裂酵母を用いた哺乳類GPCRの機能的発現の試み (2011)
  • Author(s): 清崎彰一, 猪飼篤, 長田俊哉
  • Organizer: 第84回日本生化学会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 20110921-20110924
• [Presentation] Application of Micro Scoop Cantilever for Cell Biophysics (2011)
  • Author(s): Takahiro Watanabe-Nakayama, Shin-ichi Machida, Masakazu Saito, Rehana Afrin, Atsushi Ikai
  • Organizer: 日本生物物理学会第49回年会
  • Place of Presentation: Himeji, Japan
  • Year and Date: 20110916-20110918
• [Presentation] Manipulation of Individual Stress Fibers and Focal Adhesions with AFM to Understand the Basic Mechanics of the Cellular Construction (2011)
  • Author(s): Atsushi Ikai, Rehana Afrin, Shinichi Machida, Takahiro Watanabe-Nakayama, Masakazu, Saito, Kikuo Kishimoto
  • Organizer: AFM for BioMed Paris 2011
  • Place of Presentation: Paris, France
  • Year and Date: 20110823-20110827
• [Presentation] Concerted Displacement of Stress Fibers in vivo under Direct Mechanical Manipulation (2011)
  • Author(s): Atsushi Ikai, Rehana Afrin, Shinichi Machida, Takahiro Watanabe-Nakayama, Masakazu, Saito, Kikuo Kishimoto
  • Organizer: BIT 1st Annual World Congress of Molecular & Cell Biology (CMCB-2011)
  • Place of Presentation: Beijing, China(invited)
  • Year and Date: 20110806-20110808
• [Presentation] Interaction Between Serum Proteins And Polymer Surface Studied By Atomic Force Microscopy (2011)
  • Author(s): Rehaha Afrin, Takahiro Watanabe-Nakayama, Shinichi Machida, Masakazu, Saito, Atsushi Ikai
  • Organizer: AFM-BioMed Conference
  • Place of Presentation: Paris, France
  • Year and Date: 20110803-20110827
• [Presentation] Linkage structures between ventral stress fibers revealed by atomic force and fluorescence microscopy (2011)
  • Author(s): Atsushi Ikai, Rehana Afrin, Shinichi Machida, Takahiro Watanabe-Nakayama, Masakazu, Saito, Kikuo Kishimoto
  • Organizer: Seeing is Believing : Imaging the Processes of Life
  • Place of Presentation: Santa Barbara, USA
  • Year and Date: 20110720-20110722
• [Presentation] Aomic Force Microscopy for Nano-medical Applications (2011)
  • Author(s): Rehana Afrin, Takahiro Watanabe-Nakayama, Shinichi Machida, Masakazu, Saito, Atsushi Ikai
  • Organizer: 2nd International NanoMedicine Conference
  • Place of Presentation: Sydney, Australia
  • Year and Date: 20110714-20110716
• [Presentation] Mechanical Interactions of Red Blood Cell Cytoskeleton and Associated Proteins (2011)
  • Author(s): Rehana Afrin, Takahiro Watanabe-Nakayama, Shinichi Machida, Masakazu, Saito, Atsushi Ikai
  • Organizer: 13th International Scanning Probe Microscopy Conference (ISPM)
  • Place of Presentation: Munich, Germany
  • Year and Date: 20110619-20110622
• [Presentation] マニピュレーション機能をもつカンチレバーの開発と1細胞研究への適用 (2011)
  • Author(s): 中山隆宏
  • Organizer: 東京大学生産技術研究所・JPK Instruments AG共同開催ワークショップ「生命科学における走査プローブ法と光ピンセット」
  • Place of Presentation: Tokyo, Japan(invited)
  • Year and Date: 2011-09-28
• [Presentation] Mechanical Analysis of the Bio Cell Subjected to AFM Loading (2011)
  • Author(s): Kikuo Kishimoto, David Lee, Yotaro Shinosawa, Kazuaki Inaba, Rehana Afrin, Atsushi Ikai
  • Organizer: International Workshop on Biomedical Engineering & Biomechanics
  • Place of Presentation: 西安,中国(invited)
  • Year and Date: 2011-08-13
• [Presentation] Integrity Assessment of Multi-Material Structures and Smart Structure Technology (2011)
  • Author(s): Kikuo Kishimoto
  • Organizer: ASPIRE Symposium
  • Place of Presentation: KAIST,韓国
  • Year and Date: 2011-07-07
• [Book] Molecular Manipulation with Atomic Force Microscopy(Eds.Anne-Sophie Duwez, Nicolas Willet) (2012)
  • Author(s): Ikai, A., Afrin, R., Watanabe-Nakayama, T., Machida, S.
  • Total Pages: 287
  • Publisher: CRC Press
• [Book] Life at the Nanoscale : Atomic Force Microscopy of Live Cells(Ed.Yves Dufrene) (2012)
  • Author(s): Ikai, A., Afrin, R., Watanabe-Nakayama, T., Machida, S.
  • Total Pages: 460
  • Publisher: Pan Stanford Publishing
• [Book] Atomic Force Microscopy in Liiquid(Eds.Arturo M.Baro, Ronald G.Reifenberger) (2012)
  • Author(s): Ikai, A., Afrin, R., Watanabe-Nakayama, T., Machida, S., Saito, M.
  • Total Pages: 402
  • Publisher: Wiley-VCH