Nano architecture of membrane cytoskeleton in native state revealed by immuno-cryo-electron microscopy

• Project/Area Number: 18370062
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biophysics
• Research Institution: Nagoya University
• Principal Investigator: USUKURA Jirou Nagoya University, エコトピア科学研究所, 教授 (30143415)
• Co-Investigator(Kenkyū-buntansha): TANAKA Nobuo 名古屋大, エコトピア科学研究所, 教授 (40126876) ; WATANABE Takashi 名古屋大学, 高等研究院, 特任講師 (10402562)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥17,840,000 (Direct Cost: ¥15,200,000、Indirect Cost: ¥2,640,000); Fiscal Year 2008: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2007: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000); Fiscal Year 2006: ¥6,400,000 (Direct Cost: ¥6,400,000)
• Keywords: 膜細胞骨格 / クライオ電顕 / アクチン線維 / 細胞膜 / 細胞骨格 / フリーズエッチング / 微小管 / 生物物理 / 電子顕微鏡 / 蛋白質 / 細胞・組織 / 裏打ち構造 / IQGAP1

Research Abstract

膜細胞骨格の主成分であるアクチン線維の空間構造が明らかにされた. アクチン線維は空間的に膜に密着するタイプと膜面と平行に走り所々で接着するタイプおよびストレスファイバーに分類でき、独特の網目構造作り運動と関与する線維は第二のものであることが分かった. また、これらを制御するアクチン結合タンパク質にも空間特異性があることが明らかとなった.

Research Products

• [Journal Article] Generation of transgenic rabbit model of retinal degeneration (2009)
  • Author(s): Kondo M, Sakai T, Komeima K, KurimotoY, Ueno S, Nishizawa Y, Usukura J, Fujikado T, Tano Y, Terasaki H.
  • Journal Title: Invest Ophthalmol Vis Sci. 50 Pages: 1371-1377
  • Peer Reviewed
• [Journal Article] Generation of transgenic rabbit model of retinal degeneration (2009)
  • Author(s): Usukura J
  • Journal Title: Invest Ophthalmol Vis Sci. 50 Pages: 1371-1377
  • Peer Reviewed
• [Journal Article] Pikachurin, a dystroglycan ligand, is essential for photoreceptor ribbon synapse formation (2009)
  • Author(s): Usukura J
  • Journal Title: Nature Neurosci. 11 Pages: 923-931
  • Peer Reviewed
• [Journal Article] Pikachurin, a dystroglycan ligand, is essential for photoreceptor ribbon synapse formation (2008)
  • Author(s): Sato S, Omori Y, Katoh K, Kondo M, Kanagawa M, Miyata K, Funabiki K, Koyasu T, Kajimura N, Miyoshi T, Sawai H, Kobayashi K, Tani A, Toda T, Usukura J, Tano Y, Fujikado T, Furukawa T.
  • Journal Title: Nature Neurosci 11 Pages: 923-931
  • Peer Reviewed
• [Journal Article] Three-dimensional molecular architecture of the plasma-membrane-associated cytoskeleton as (2008)
  • Author(s): Morone N, Nakada C, Umemura Y, Usukura J, Kusumi A
  • Journal Title: reconstructed by freeze-etch electron tomography Methods Cell Biol 88 Pages: 207-236
  • Peer Reviewed
• [Journal Article] Ultrastructural study of Rac1 and its effectors beneath the substratum-facing membrane (2008)
  • Author(s): Watanabe T, Wang S, Kakeno M, Usukura J, Kaibuchi K.
  • Journal Title: Cell Struct Funct 33 Pages: 101-107
  • NAID: 130004137531
  • Peer Reviewed
• [Journal Article] Gating-associated conformational changes in the mechanosensitive channel MscL (2008)
  • Author(s): Yoshimura K, Usukura J, Sokabe M.
  • Journal Title: Proc Natl Acad Sci U S A. 105 Pages: 4033-8
  • Peer Reviewed
• [Journal Article] Regulation of VEGF-mediated angiogenesis by the Akt/PKB substrate Girdin (2008)
  • Author(s): Kitamura T, Asai N, Enomoto A, Maeda K, Kato T, Ishida M, Jiang P, Watanabe T, Usukura J, Kondo T, Constantini F, Murohara T, Takahashi M.
  • Journal Title: Nature Cell Biol 10 Pages: 329-337
  • Peer Reviewed
• [Journal Article] Ultrastructural study of Racl and its effectors beneath the substratum-facing membrane. (2008)
  • Author(s): Usukura J
  • Journal Title: Cell struct Funct. 33 Pages: 101-107
  • Peer Reviewed
• [Journal Article] Three-dimensional molecular architecture of the plasma-membrane-associated cytoskeleton as reconstructed by freeze-etchelectron tomography (2008)
  • Author(s): Usukura J
  • Journal Title: Methods Cell Biol. 88 Pages: 207-236
  • Peer Reviewed
• [Journal Article] Characterizaion of native myosin VI isolated from sea urchin eggs (2007)
  • Author(s): Sakata S, Watanabe Y. Usukura J, Mabuchi I.
  • Journal Title: J Biochem 42 Pages: 481-90
  • Peer Reviewed
• [Journal Article] Physiological properties of rod photoreceptor cells in green-sensitive cone pigment knock-in mice (2007)
  • Author(s): Sakurai K, Onishi A, Imai H, Chisaka O, Ueda Y, Usukura J, Nakatani K, Shichida Y.
  • Journal Title: J Gen Physiol 130 Pages: 21-40
  • Peer Reviewed
• [Journal Article] LEDGF/DFS70, a mojor autoantigen of atopic dermatitis, is a component of keratohyalin granules (2007)
  • Author(s): Sugiura K, Muro Y, Nishizawa Y,Okamoto M, Shinohara T, Tomita Y, Usukura J.
  • Journal Title: J. Invest. Dermatol 127 Pages: 75-80
• [Journal Article] Characterization of native myosin VI isolated from sea urchin eggs (2007)
  • Author(s): Usukura J
  • Journal Title: J Biochem. 42 Pages: 481-490
  • NAID: 10020096069
  • Peer Reviewed
• [Journal Article] Physiological properties of rod photoreceptor cells in green-sensitivecone pigment knock-in mice (2007)
  • Author(s): Usukura J
  • Journal Title: J Gen Physiol. 130 Pages: 21-40
  • Peer Reviewed
• [Journal Article] Three-dimensional reconstruction of the membrane skeleton at the plasma membrane interface by electron tomography (2007)
  • Author(s): Usukura J
  • Journal Title: J. Cell Biol. 174 Pages: 851-862
  • Peer Reviewed
• [Journal Article] LEDGF/DFS70, a mojor autoantigen of atopic dermatitis, is a component of keratohyalin granules. (2007)
  • Author(s): Usukura J
  • Journal Title: J. Invest. Dermatol. 127 Pages: 75-80
• [Journal Article] Three-dimensional reconstruction of the membrane skeleton at the plasma membrane interface by electron tomography (2006)
  • Author(s): Morone N, Fujiwara T, Murase K, Kasai S, Ike H, Yuasa S, Usukura J, Kusumi A.
  • Journal Title: J. Cell Biol 174 Pages: 851-862
  • Peer Reviewed
• [Journal Article] Structural insight of human DEAD-box protein rck/p54 into its substrate recognition with conformational changes (2006)
  • Author(s): Matsui T, Hogetsu K, Usukura J, Sato T, Kumasaka T, Akao Y, Tanaka N.
  • Journal Title: Genes Cells 11 Pages: 439-452
  • Peer Reviewed
• [Journal Article] Structural insight of human DEAD-box protein rck/p54 into its substrate recognition with conformational changes (2006)
  • Author(s): Usukura J
  • Journal Title: Genes Cells 11 Pages: 439-452
• [Journal Article] Illuminated rhodopsin is required for strong activation of retinal guanylate cyclase by guanylate cyclase-activating proteins. (2006)
  • Author(s): Usukura J
  • Journal Title: Biochemistry 45 Pages: 1899-1909
• [Presentation] 3D architecture of membrane cytoskeleton and spatial specificity of actin binding proteins revealed by immuno-freeze-etching electron microscopy (2009)
  • Author(s): Usukura J
  • Organizer: Ninth International Symposium on Biomimetic Materials Processing January
  • Place of Presentation: Nagoya Japan
• [Presentation] 3D architecture of membrane cytoskeleton and spatial specificity of actin binding proteins revealedby immuno-freeze-etching electron microscopy. (2009)
  • Author(s): Usukura J
  • Organizer: Ninth International Symposium on Biomimetic Materials Processing
  • Place of Presentation: Nagoya Japan
• [Presentation] Molecular shapes reconstructed on the artificial membrane. The First iCeMS International Symposium The Eleventh (2008)
  • Author(s): 臼倉治郎
  • Organizer: MEMBRANE RESEARCH FORUM
  • Place of Presentation: 京都フジタホテル
  • Year and Date: 2008-02-21
• [Presentation] Spatial archite cture of membrane cytoskeleton and spatial specificity of actin binding proteins revealed by immuno- freeze-etching and cryo-microscopy (2008)
  • Author(s): Usukura J.
  • Organizer: The 48th Annual meeting of the American Society for Cell Biology
  • Place of Presentation: San Francisco, USA
• [Presentation] 3D Architecture of Membrane Cytoskeleton and Spatial Specificity of Actin Binding Proteins Revealed by Immuno-freeze- etching and Cryo-microscopy (2008)
  • Author(s): Usukura J.
  • Organizer: The 39 NIPS International Symposium
  • Place of Presentation: Okazaki Japan
• [Presentation] 3D Architecture of Membrane of Undercoat and Spatial Specificity of Actin Binding Protein Localization (2008)
  • Author(s): Usukura J.
  • Organizer: The 5th International Form on Oxidative Stress and Aging
  • Place of Presentation: Ancona, Italy
• [Presentation] 3D architecture of membrane undercoat and spatial specificity of actin binding proteins revealed by immuno-freeze-etching andcryo-electron microscopy. XIII Interntional Congress of Histochemistry and Cytochemistry (2008)
  • Author(s): Usukura J.
  • Organizer: Congress of Histochemistry and Cytochemistry
  • Place of Presentation: Medical University of Gdansk, Poland
• [Presentation] A Pilot Study for the Assessment on a Biological Toxicity Caused by Nano-particles (2008)
  • Author(s): 臼倉治郎
  • Organizer: Symposium on Biomimetic Materials Processing
  • Place of Presentation: Noyori Conference Hall
• [Presentation] 3D architecture of membrane undercoat and spatial specificity of actin binding proteins revealed by immuno-freeze-etching and cryo-electron microscopy (2008)
  • Author(s): Usukura J
  • Organizer: XIII International Congress of Histochemistry and Cytochemistry.
  • Place of Presentation: Medical University of Gdansk, Poland
• [Presentation] 3D Architecture of Membrane of Undercoat and Spatial Specificity of Actin Binding Protein Localization (2008)
  • Author(s): Usukura J
  • Organizer: The 5^<th>International Forum on Oxidative Stressand Aging
  • Place of Presentation: Ancona, Italy
• [Presentation] 3D Architecture of Membrane Cytoskeleton and Spatial Specificity of Actin Binding Proteins Revealed by Immuno-freeze-etching and Cryo-microscopy. (2008)
  • Author(s): Usukura J
  • Organizer: The 39 NIPS International Symposium
  • Place of Presentation: Okazaki Japan
• [Presentation] Spatial architecture of membrane cytoskeleton and spatial specificity of actin binding proteins revealed by immuno-freeze-etching and cryo-microscopy. (2008)
  • Author(s): Usukura J
  • Organizer: 48^<th> Annual meeting of the American Societyfor Cell Biology
  • Place of Presentation: San Francisco, USA
• [Presentation] 次世代電子顕微鏡技術による細胞内空間構造の解明を目指して (2007)
  • Author(s): 臼倉治郎
  • Organizer: 日本電子顕微鏡学会
  • Place of Presentation: 新潟コンベンションセンター
  • Year and Date: 2007-05-21
• [Presentation] Cryo-electron microscopic analysis of three dimensional membrane cytoskeleton (2007)
  • Author(s): 臼倉治郎
  • Organizer: 第40回日本発生生物学会、第59回日本細胞生物学会合同大会
  • Place of Presentation: 福岡国際会議場
• [Book] よくわかる生物電子顕微鏡技術-プロトコール・ノウハウ・原理 (2008)
  • Author(s): 臼倉治郎
  • Total Pages: 200
  • Publisher: 共立出版
• [Book] よくわかる生物電子顕微鏡技術-プロとコール・ノウハウ・一原理- (2008)
  • Author(s): 臼倉治郎
  • Total Pages: 200
  • Publisher: 共立出版(株)
• [Remarks]
  • URL: http://www.numse.nagoya-u.ac.jp/bio
• [Remarks]
  • URL: http://www.numse.nagoya-u.ac.jp/bio