Structural analyses of the interaction between extracellular ligands and their receptors

2009 Fiscal Year Final Research Report

• Project/Area Number: 17082004
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: TAKAGI Junichi Osaka University, 蛋白質研究所, 教授 (90212000)
• Co-Investigator(Kenkyū-buntansha): IWASAKI Kenji 大阪大学, 蛋白質研究所, 准教授 (20342751)
• Co-Investigator(Renkei-kenkyūsha): NOGI Terukazu 大阪大学, 蛋白質研究所, 助教 (40379102) ; YASUI Norihisa 大阪大学, 蛋白質研究所, 助教 (90467514)
• Project Period (FY): 2005 – 2009
• Keywords: シグナル伝達 / 構造解析 / リガンド複合体

Research Abstract

We have utilized X-ray crystallography and electron microscopy to decipher the structural mechanism underlying recognition of extracellular environmental cues by cellular receptors. We have gained information about reelin signal transduction in brain development and Wnt signal transduction implicated in cancer and bone formation. Also, we have unraveled the structure of neuronal synapse adhesion machinery, and succeeded in visualizing the rice dwarf virus during the infection in situ.

Research Products

• [Journal Article] Structural basis for specific recognition of reelin by its receptors. (2010)
  • Author(s): Yasui N, Nogi T, Takagi
  • Journal Title: Structure 18 Pages: 320-331
  • Peer Reviewed
• [Journal Article] Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation. (2010)
  • Author(s): Yasui N, Mihara E, Nampo M, Tamura-Kawakami K, Unno H, Matsumoto K, Takagi
  • Journal Title: J. immunol. Methods 352 Pages: 153-160
  • Peer Reviewed
• [Journal Article] Structural insights into von Willebrand factor cleavage by ADAMTS13. (2009)
  • Author(s): Akiyama, M., Takeda, S., Kokame, K., L Takagi, Miyata, T.
  • Journal Title: Proc. Natl. Acad. Sci. USA (in press)
  • Peer Reviewed
• [Journal Article] Production, crystallization and preliminary crystallographic analysis of the exosite-containing fragment of human von Willebrand factor-cleaving proteinase, ADAMTS 13. (2009)
  • Author(s): Akiyama, M., Takeda, S., Kokame, K., L Takagi., Miyata, T.
  • Journal Title: Acta Crystallographica. F65 Pages: 739-742
  • Peer Reviewed
• [Journal Article] Association of Rice gall dwarf virus with microtubules is necessary for viral release from cultured insect vector cells (2009)
  • Author(s): T. Wei, T. Ichiki-Uehara, N. Miyazaki, H. Hibino, K. Iwasaki, T. Omura
  • Journal Title: J. Virol. 83(20) Pages: 10830-10835
  • Peer Reviewed
• [Journal Article] A self-assembled protein nanotubule with high aspect ratio (2009)
  • Author(s): Frederico F. Miranda, Kenji Iwasaki, Satoko, Akashi, Koji Sumitomo, Mime Kobayashi, Ichiro Yamashita, Jeremy R.H. Tamae, Jonathan G. Heddle.
  • Journal Title: Small. 5 Pages: 2077-2084
  • Peer Reviewed
• [Journal Article] The structure of rat liver vault at 3. 5 angstrom resolution, H. Tanaka, K. Kato, E. Yamashita (2009)
  • Author(s): H. Tanaka, K. Kato, E. Yamashita, T. Sumizawa, Y. Zhou, M. Yao K. Iwasaki, M. Yoshimura, T. Tsukihara
  • Journal Title: Science 323 Pages: 384-388
  • Peer Reviewed
• [Journal Article] Molecular basis of the recognition of nephronectin by integrin alpha8betal. (2009)
  • Author(s): Sato, Y., Uemura, T., Morimitsu, K., Sato-Nishiuchi, R., Manabe, R., Takagi, J., Yamada, M., Sekiguchi, K.
  • Journal Title: J. Biol. Chem. 284 Pages: 14524-14536
  • Peer Reviewed
• [Journal Article] Structure of F-spondin reeler domain reveals a unique β-sandwich fold with a deformable disulfide-bonded loop. (2008)
  • Author(s): Nagae, M., Nishikawa, K., Yasui, N., Yamasaki, M., Nogi, T., Takagi, J.
  • Journal Title: Acta Crystallographica D64 Pages: 1138-1145
  • Peer Reviewed
• [Journal Article] Novel affinity tag system using structurally defined antibody-tag interaction: Application to single-step protein purification. (2008)
  • Author(s): Nogi, T., Sangawa, T., Tabata, S., Nagae, M., Tamura-Kawakami, K., Beppu, A., Hattori, M., Yasui, N., Takagi, J.
  • Journal Title: Protein Sci. 17 Pages: 2120-2126
  • Peer Reviewed
• [Journal Article] A murine monoclonal antibody that binds N-terminal extracellular segment of human protease activated receptor-4. (2008)
  • Author(s): Sangawa, T., Nogi, T., Takagi, J.
  • Journal Title: Hybridoma 27(5) Pages: 331-335
  • Peer Reviewed
• [Journal Article] Key interactions in ectodomain of integrin for global conformational change detected by elastic network normal mode analysis. (2008)
  • Author(s): Atsushi Matsumoto, Tetsuji Kamata, Junichi Takagi, Kenji Iwasaki, Kei Yura.
  • Journal Title: Biophys. J. 95(6) Pages: 2895-2908
  • Peer Reviewed
• [Journal Article] Milk fat globule EGF factor 8 in the serum of human patients of systemic lupus erythematosus. (2008)
  • Author(s): Yamaguchi, H., Takagi, J., Miyamae, T., Yokota, S., Fujimoto, T., Nakamura, S., Ohshima, S., Naka, T., Nagata, S.
  • Journal Title: J. Leukocyte Biol. 83 Pages: 1300-1307
  • Peer Reviewed
• [Journal Article] Structural basis for ligand recognition by integrins. (2007)
  • Author(s): Takagi, J.
  • Journal Title: Curr. Opin. Cell Biol. 19 Pages: 557-564
• [Journal Article] receptor-binding reelin fragment and mutational analysis reveal a recognition mechanism similar to endocytic receptors. (2007)
  • Author(s): Yasui, N., Nogi, T., Kitao, T., Nakano, Y., Hattori, M., Takagi, J.
  • Journal Title: Proc. Natl. Acad. Sci. USA 104 Pages: 9988-9993
  • Peer Reviewed
• [Journal Article] The RGD motif in fibronectin is essential for mouse development but dispensable for fibronectin fibril assembly. (2007)
  • Author(s): Takahashi, S., Leiss, M., Moser, M., Ohashi, T., Kitao, T., Heckmann, D., Pfeifer, A., Kessler, H., Takagi, T., Erickson, H.P., Fassler, R.
  • Journal Title: J. Cell Biol. 178 Pages: 167-178
  • Peer Reviewed
• [Journal Article] Three-dimensional architecture of virus-packed tubule. (2007)
  • Author(s): Katayama, S., Wei, T., Omura, T., Takagi., Iwasaki, K.
  • Journal Title: J. Electron Microsc. 56 Pages: 77-81
  • Peer Reviewed
• [Journal Article] Semaphorin 7A initiates T cell-mediated inflammatory responses through a1131 integrin. (2007)
  • Author(s): Suzuki K., Okuno, T., Yamamoto, M., Pasterkamp, R.J., Takegahara, N., Kitao, T., Takagi, J., Rennert, P.D., Kolodkin, A.L., Kumanogoh, A., Kikutani, H.
  • Journal Title: Nature 446 Pages: 680-684
  • Peer Reviewed
• [Journal Article] aVr36 is a novel receptor for human fibrillin-1: comparative studies of molecular determinants underlying integrin-RGD affinity and specificity. (2007)
  • Author(s): Jovanovic J, Takagi J, Choulier L, Abrescia NG, Stuart DI, van der Merwe PA, Mardon HJ, Handford PA
  • Journal Title: J. Biol. Chem. 282 Pages: 6743-6751
  • Peer Reviewed
• [Journal Article] Structure of a signaling-competent reelin fragment revealed by X-ray crystallography and electron tomography. (2006)
  • Author(s): Nogi, T., Yasui, N., Hattori, M., Iwasaki, K., Takagi, J.
  • Journal Title: EMBO J. 25 Pages: 3675-3683
  • Peer Reviewed
• [Journal Article] モジュラータンパク質の構造解析:受容体構造生物学における中心課題 (2006)
  • Author(s): 高木淳一
  • Journal Title: 日本結晶学会誌 48 Pages: 411-417
• [Journal Article] Disrupting integrin transmembrane domain heterodimerization increases ligand binding affinity, not valency or clustering. (2005)
  • Author(s): Luo, B.H., Carman, C.V., Takagi, J., Springer, T.A.
  • Journal Title: Proc Natl Acad Sci U S A 102 Pages: 3679-3684
  • Peer Reviewed
• [Journal Article] Potentiation of the ligand-binding activity of integrin alpha3 betal via association with tetraspanin CD151. (2005)
  • Author(s): Nishiuchi, R., Sanzen, N., Nada, S., Sumida, Y., Wada, Y., Okada, M., Takagi, J., Hasegawa, H., Sekiguchi, K.
  • Journal Title: Proc Natl Acad Sci U S A 102 Pages: 1939-1944
  • Peer Reviewed
• [Journal Article] 電子顕微鏡を用いた蛋白質の高次構造解析が開く世界-インテグリンレセプターの構造解析を中心として- (2005)
  • Author(s): 岩崎憲治、高木淳一
  • Journal Title: 生化学 77 Pages: 400-410
• [Presentation] 神経シナプスの形成を制御するニューレキシン・ニューロリギン複合体の結晶構造 (2010)
  • Author(s): 禾晃和
  • Organizer: 第23回日本放射光学会年会放射光科学合同シンポジウム
  • Place of Presentation: 姫路
  • Year and Date: 2010-01-08
• [Presentation] EM imaging of Rice Dwarf Virus (2010)
  • Author(s): Iwasaki, K.
  • Organizer: China-Japan 3D-EM Forum
  • Place of Presentation: Beijing(中国)
  • Year and Date: 2010-01-05
• [Presentation] シナプス間接着分子Neuroligin/Neurexin複合体の構造から見えるシナプス間隙の真の姿 (2009)
  • Author(s): 高木淳一
  • Organizer: 第5回プロテオミクス・構造生物学講演会
  • Place of Presentation: 東京
  • Year and Date: 2009-11-22
• [Presentation] リーリン会合体の構造的基盤とその機能的意義 (2009)
  • Author(s): 安井典久
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2009-10-24
• [Presentation] Dissection of the molecular architecture of higher-order assembly of the full-length reelin (2009)
  • Author(s): Yasui, N.
  • Organizer: 23rd Symposium of The Protein Society
  • Place of Presentation: Boston(USA)
  • Year and Date: 2009-07-28
• [Presentation] Crystal structure of synaptic adhesion complex; insights into the molecular assembly at the synaptic membrane (2009)
  • Author(s): Takagi, J.
  • Organizer: 23rd Symposium of The Protein Society
  • Place of Presentation: Boston(USA)
  • Year and Date: 2009-07-27
• [Presentation] Electron microscopy study of platelet and integrin using new methods (2009)
  • Author(s): Iwasaki, K.
  • Organizer: 第32回日本血栓止血学会学術集会
  • Place of Presentation: 小倉
  • Year and Date: 2009-06-04
• [Presentation] CEMOVISの評価とクライオ電子線トモグラフィー (2009)
  • Author(s): 岩崎憲治
  • Organizer: 日本顕微鏡学会第65回学術講演会
  • Place of Presentation: 仙台
  • Year and Date: 2009-05-27
• [Presentation] 全長リーリンの精製法の確立とリーリン会合体の構造化学的基盤の解明 (2009)
  • Author(s): 安井典久
  • Organizer: 第9回日本蛋白質科学会年会
  • Place of Presentation: 熊本
  • Year and Date: 2009-05-22
• [Presentation] クライオ電子顕微鏡法を用いた活性化血小板の分子分解能構造解析 (2008)
  • Author(s): 片山寿美枝
  • Organizer: 第81回日本生化学会大会
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-10
• [Presentation] Cryo-Electron Tomography for Single Particles (2008)
  • Author(s): Iwasaki, K.
  • Organizer: 9th Asia-Pacific Microscopy Conference
  • Place of Presentation: Jeju (Korea)
  • Year and Date: 2008-11-02
• [Presentation] Crystal structure ofsynaptic adhesion protein neurexin and neuroligin (2008)
  • Author(s): Tanaka, H.
  • Organizer: XXI Congress of the International Union of Crystallography
  • Place of Presentation: Osaka
  • Year and Date: 2008-08-23
• [Presentation] Structural basis for recognition of reelin by its neuronal receptor (2008)
  • Author(s): Yasui, N.
  • Organizer: 22nd Annual Symposium of The Protein Society
  • Place of Presentation: San Diego (USA)
  • Year and Date: 2008-07-29
• [Presentation] リーリン工受容体複合体の結晶構造解析 (2008)
  • Author(s): 安井典久
  • Organizer: 第8回日本蛋白質科学会年会
  • Place of Presentation: 東京
  • Year and Date: 2008-06-10
• [Presentation] F-spondinリーラードメインのX線結晶構造解析 (2008)
  • Author(s): 長江雅倫
  • Organizer: 第8回日本蛋白質科学会年会
  • Place of Presentation: 東京
  • Year and Date: 2008-06-10
• [Presentation] 脳の層構造形成を司る細胞外因子リーリンの結品構造と受容体による認識機構 (2007)
  • Author(s): 高木淳一
  • Organizer: 日本分子生物学会・日本生化学会合同シンポジウム(BMB2007)
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-15
• [Presentation] シナプス間接着タンパク質αニューレキシンの細胞外ドメインの構造解析 (2007)
  • Author(s): 田中宏樹
  • Organizer: 日本結晶学会2007年度年会
  • Place of Presentation: 東京
  • Year and Date: 2007-12-02
• [Presentation] Challenges in structural analysis of extracellular protein complexes (2007)
  • Author(s): Takagi.
  • Organizer: 2nd International Symposium on Diffraction Structural Biology 2007
  • Place of Presentation: Tokyo
  • Year and Date: 2007-09-11
• [Presentation] Crystal Structure of a Signaling- competent Reelin Fragment (2007)
  • Author(s): Nogi, T.
  • Organizer: The 2007 Meeting of the American Crystallographic Association
  • Place of Presentation: Salt Lake City(USA)
  • Year and Date: 2007-07-25
• [Presentation] 受容体によるリーリン認識の構造的基盤 (2007)
  • Author(s): 安井典久
  • Organizer: 第7回日本蛋白質科学会年会
  • Place of Presentation: 仙台
  • Year and Date: 2007-05-25
• [Book] インテグリン、「ますます重要になる細胞周辺環境(細胞ニッチ)の最新科学技術」(田畑泰彦編) (2009)
  • Author(s): 高木淳一
  • Total Pages: 251-256
  • Publisher: ディカルドゥ
• [Book] Reelin Glycoprotein, biology, structure and roles in health and disease (2008)
  • Author(s): Takagi, J.
  • Total Pages: 1-444
  • Publisher: Springer
• [Book] ポストゲノム時代のタンパク質科学-構造・機能・ゲノミクス- (Arthur M. Lesk原著) (2007)
  • Author(s): 高木淳一
  • Total Pages: 1-328
  • Publisher: 化学同人
• [Book] 再生医療のための細胞生物学(関口清俊編) (2007)
  • Author(s): 高木淳一
  • Total Pages: 26-48
  • Publisher: コロナ社
• [Remarks]
  • URL: http://www.protein.osaka-u.ac.jp/rcsfp/synthesis/index.html
• [Patent(Industrial Property Rights)] プロテアーゼ認識配列を有するタグペプチドおよびその利用 (2009)
  • Inventor(s): 高木淳一
  • Industrial Property Rights Holder: 国立大学法人大阪大学
  • Industrial Property Number: 特許・特願2009-103925
  • Filing Date: 2009-04-22
• [Patent(Industrial Property Rights)] タグペプチド及びその利用 (2008)
  • Inventor(s): 高木淳一
  • Industrial Property Rights Holder: 国立大学法人大阪大学
  • Industrial Property Number: 特許・PCT/JP2008/073069
  • Filing Date: 2008-12-18
  • Overseas
• [Patent(Industrial Property Rights)] タグペプチドが結合した融合タンパク質、タグペプチドに対する抗体及びこれらを用いるタンパク質の精製方法 (2008)
  • Inventor(s): 高木淳一
  • Industrial Property Rights Holder: 国立大学法人大阪大学
  • Industrial Property Number: 特許・特願2008-020804
  • Filing Date: 2008-01-31