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Role of three dimensional structure of proteins in signal transduction

Research Project

Project/Area Number 06276105
Research Category

Grant-in-Aid for Scientific Research on Priority Areas

Allocation TypeSingle-year Grants
Research InstitutionTokyo Metropolitan Organization for Medical Research

Principal Investigator

INAGAKI Fuyuhiko  Tokyo Metropolitan Institute of Medical Science, Researcher, 生理活性物質研究部門, 研究員 (70011757)

Co-Investigator(Kenkyū-buntansha) MIZUNO Takeshi  Nagoya Univ., School of Agriculture, Professor, 農学部, 教授 (10174038)
YAMAMOTO Tadashi  Tokyo Univ., Institute of Medical Science, Professor, 医科学研究所, 教授 (40134621)
TAKENAWA Tadaomi  Tokyo Univ., Institute of Medical Science, Professor, 医科学研究所, 教授 (40101315)
OONO Shigeo  Yokohama City Univ., School of Medicine, Professor, 医学部, 教授 (10142027)
KOHDA Daisuke  Biomolecular Engineering Research Institute, 研究員 (80186618)
伊倉 光彦  筑波大学, 先端学際領域センター, 教授 (00142688)
今川 正良  大阪大学, 薬学部, 助教授 (20136823)
片平 正人  横浜国立大学, 工学部, 講師 (70211844)
Project Period (FY) 1994 – 1997
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥106,300,000 (Direct Cost: ¥106,300,000)
Fiscal Year 1997: ¥17,300,000 (Direct Cost: ¥17,300,000)
Fiscal Year 1996: ¥24,500,000 (Direct Cost: ¥24,500,000)
Fiscal Year 1995: ¥31,500,000 (Direct Cost: ¥31,500,000)
Fiscal Year 1994: ¥33,000,000 (Direct Cost: ¥33,000,000)
Keywordsthree dimensional structure of protein / NMR / X-ray crystallography / Grb2 / SH2 / SH3 / destrin / midkine / Ash / 立体構造 / N-WASP / フィロポディア / 二成分系 / リン酸基転移反応 / 嫌気センサー / GRB2 / 神経突起伸長 / NMR / ArcB / Tob / リン酸化チロシン / プロリンに富むペプチド / 溶液構造 / Ras / ポリプロリンヘリックス / Solution Structure
Research Abstract

The aim of the present research was to elucidate the roles of proteins in signal transduction process on the structural basis. We also developed the protein expression and purification systems for preparation of amount of proteins required for structural studies. In the course of the present research project, a number of protein structures were determined by either NMR spectroscopy or X-ray crystallography. Grb2 n-SH3 and SH2 domains complexed with proline rich peptide from Sos and phosphotyrosine containing peptide from Shc, respectively were determined and the general mechanism for recognition of target sequences were elucidated. Three dimensional structure of destrin was determined by NMR. Although the sequence homology between destrin family and gelsolin family proteins was not present, the three dimensional structure of both proteins were very similar, suggesting the same mechanism for actin severing. Thus, the general mechanism for severing of the actin fiber was proposed. The three dimensional structure of midkine was also determined by NMR and elucidated the mechanism of dimer formation on heparin oligosaccharides. The dimer formation of midkine seems to be essential for activation of the receptor and its biological function. We also determined the CRD domain of PKC and suggested the mechanism of DAG recognition. By X-ray crystallography, a number of protein structures were determined including transmitter and receiver domains of bacterial signal transduction system. We also made protein expression and purification system for N-WASP, Tob and CAF1. N-WASP is essential for formation of philopodia in response to the growth factor receptor signal. Tob and CAF1 play pivotal role in cell cycling. The structural studies started for both proteins.

Report

(5 results)
  • 1998 Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • 1994 Annual Research Report
  • Research Products

    (46 results)

All Other

All Publications (46 results)

  • [Publications] Kohda,D.: "Solution structure and ligand-binding site of the carboxy-terminal SH3 domain of GRB2."Structure. 2. 1029-1040 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Terasawa,H.: "Structure of the N-terminal SH3 domain of GRB2 complexed with a peptide from the guanine nucleotide releasing factor Sos"Nature structural biology. 1. 891-897 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Iwasaki,W.: "Solution structure midkine,a new heparin-binding growth factor."ENBO J.. 16. 6936-6946 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kohda,D.: "olusion Structure of the Link Module:A Hyaluronan-Binding Domain Involved in Extracellular Matrix Stability and Cell Migration."Cell. 86. 767-775 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hatanaka,H.: "Tertiary Structure of Dastrin and Structural Similarity between Two Actin-Regulating Protein Families."Cell. 85. 1047-1055 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ogura,K.: "An impruved double-tuned and isotope-filtered pulse scheme based on a pulsed field gradient and a wide-band inversion shaped pulse."J.Biomol.NMR. 8. 492-498 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 稲垣冬彦: "SH3によるプロチンに富む配列の認識(構造生物学とその解析法)"共立出版(京極好正、月原富武編). 7(192) (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kohda, D., Terasawa, H., Ichikawa, S., Ogura, K., Hatanaka, H., Mandiyan, V., Ullrich, A., Schlessinger, J. and Inagaki, F.: "Solution structure and ligand-binding site of the carboxy-terminal SH3 domain of GRB2"Structure. 2. 1029-1040 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Terasawa, H., Kohda, D., Hatanaka. H., Tsuchiya, S., Ogura, K., Nagata, K., Ishii, S., Mandiyan, V., Ullrich, A., Schlessinger, J. and Inagaki, F.: "Structure of the N-terminal SH3 domain of GRB2 complexed with a peptide from the guanine nucleotide releasing factor Sos"Nature structural biology. 1. 891-897 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hatanaka, H., Ogura, K., Moriyama, K., Ichikawa, S., Yahara, I. and Inagaki, F.: "Tertiary Structure of Destrin and Structural Similarity between Two Actin-Regulating Protein Families"Cell. 85. 1047-1055 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kohda, D., Morton, C. J., Parker, A. A., Hatanaka, H., Inagaki, F., Campbell, I. D. and Day, A. J.: "Solution Structure of the Link Module : A Hyaluronan-Binding Domain Involved in Extracellular Matrix Stability and Cell Migration"Cell. 86. 767-775 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ogura, K., Terasawa, H. and Inagaki, F.: "An improved double-tuned and isotope-filtered pulse scheme based on a pulsed field gradient and a wide-band inversion shaped pulse"J. Biomokecular NMR. 8. 492-498 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Iwasaki, W., Nagata, K., Hatanaka, H., Inui, T., Kimura, T., Muramatsu, T., Yoshida, K., Tasumi, M. and Inagaki, F.: "Solution structure of midkine, a new heparin-binding growth factor"EMBO J.. 16. 6936-6946 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sakisaka, T., Ito, T., Miura. K. and Takenawa, T.: "Phosphatidylinositol 4,5-bisphosphate phosphatase regulates the rearrangement of actin filaments"Mol. Cell. Biol.. 17. 3641-3649 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Umemori, H., Inoue, T., Kume, S., Sekiyama, N., Nagao, M., Itoh, H., Nakanishi, S., Mikoshiba, K. and Yamamoto, T: "Activation of the G protein Gq/11 through turosine phosphorylation of the a subunit"Science. 276. 1878-1881 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasue, T., Nishizumi, H., Miyake, K., Aizawa, S., Yamamoto, T., Uehara, S., Kikuchi, Y. and Takatsu, K.: "IgG1 production induced by CD38 ligation and IL-5 require activation of Fyn and Lyn kinases"Proc. Natl. Acad. Sci. USA94. 10307-10312 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Lu, Z., Hornia, A., Jiang, Y-W., Frankel, P., Zang Q., Ohno, S. and Foster, D., A.: "Tumor-promotion by deple\\ting cells of protein kinase Cd."Molecular and Cellular Biol.. 17. 3418-3428 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kato, M., Mizuno, T., Shimizu, T. and Hakoshima, T.: "Insights into multistep phosphorelay from the crystal structure of the C-terminal HPt domain of ArcB"Cell. 88. 717-723 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ames, J. B., Ishima, R., Tanaka, T., Gordon, J. I., Stryer, L. and Ikura, M.: "Molecular mechanics of calcium-myristoyl switches"Nature. 389. 198-202 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Iwasaki,W.: "Solution structure of midkine,a new heparin-binding growth factor." EMBO J.16. 6936-6946 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ogura,K.: "Comformation of an Shc-derived phosphotyrosin-containing peptide complexed with the Grb2 SH2 domain." J.bilmol.NMR. 10. 273-278 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Miki,H.: "Induction of filopodium formation by a WASP-related actin depolymerizing protein N-WASP." Nature. 391. 93-98 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Sakisaka,T.: "Phosphatidylinositol 4,5-bisphosphate phosphatase regulates the rearrangement of actin filaments." Mol.Cell Biol.17. 3841-3849 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yoshida,Y.: "Cloning and characterization of the mouse Tob Gene." Gene. 191. 109-113 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato,M.: "Insights into multistep phosphorelay from the crystal structure of the C-terminal HPt domain of ArcB." Cell. 88. 717-723 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 稲垣冬彦: "SH3によるプロリンに富む配列の認識(構造生物学とその解析法)" 共立出版(京極好正、月原富武編), 7(192) (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 稲垣冬彦: "磁気分光法(分析計測法)" 丸善(猪飼篤編), 30(187) (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Ogura,K.: "An improved double-tuned and isotope-filtered pulse scheme based on a pulsed field gradient and a wide-band inversion shaped pulse." J.Biomolecular NMR. 8. 492-498 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kohda,D.: "Solution Structure of the Link Module:A Hyaluronan-Binding Domain Involved in Extracellular Matrix Stability and Cell Migration." Cell. 86. 767-775 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Hatanaka,H.: "Tertiary Structure of Destrin and Structural Similarity between Two Actin-Regulating Protein Families." Cell. 85. 1047-1055 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ogura,K.: "^<13>C Fully Refocused Multidimensional ^<13>C-Edited Pulse Schemes Using Broadband Shaped Inversion and Refocusing Pulses." J.Magn.Reson.B112. 63-68 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Miki,H.: "A novel actin-depolymerizing protein N-WASP regulates the cortical cytoskeletal rearrangement in a PIP2 dependent manner at the downstream of tyrosine kinase." EMBO J.15. 5326-5335 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ishida,T.: "TRAF5,a novel tumor necrosis factor receptor-associated factor family protein,mediates CD40 signaling." Proc.Natl.Acad.Sci.USA.93. 9437-9442 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Fujimoto,J: "Characterization of the transforming activity of p80,a hyperphosphorylated protein in a Ki-1 lymphoma cell line with t(2;5)." Proc.Natl.Acad.Sci.USA.93. 4181-4186 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Ichikawa, S.: "Solution Structure of Cysteine-Rich Domain of Protein Kinase Cx." Journal of Biochemistry. 117. 566-574 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kohda, D.: "Trajectory analysis of NMR structure calculations." Journal of Biomolecular NMR. 5. 357-361 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nagata, K.: "Three-dimensional Solution Structure of Bombyxin-II an Insulin-like Peptide of the Silkmoth Bombyx mori : Structural Comparison with Insulin and Relaxin." J. Molecular Biology. 253. 749-758 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nagata, K.: "Identification of the Receptor-recognition Surface of Bombyxin-II, an Insulin-like Peptide of the Silkmeth Bombxy mori : Critical Importance of the B-chain Centnal Part." J. Molecular Biology. 253. 759-770 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Hashimoto, R.: "N-terminal Deletion Mutants of Insulin-like Growth Factor-II (IGF-II) Show Thr^7 and Leu^8 Important for Binding to Insulin and IGF-I Receptors and Leu^8 Critical for All IGF-II Functions." The Journal of Biological Chemistry. 270. 18013-18018 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Ogra, K.: "^<13>C Fully Refocused Multi-dimensional ^<13>CEdited Pulse Sohemes Using Broadband Shaped Inversion and Refocusing Pulses." Journal of Magnetic Resonance. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] Nagata,K.: "Solution Structure of the epiclermal growth factor like domain of heregulin-a,aligand for pl80^<erb4>." The EMBO Journal. 13. 3517-3523 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kohda,D.: "Solution Structure of human insulin-like growth factor II;recognition sites for receptors and binding proteins." Structure. 2. 1029-1040 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Terasawa,H.: "Structure of the N-terminal SH3 domain of GRB2 complexed with a peptide from the guanine nucleotide releasing factor Sos." Nature structual biology.1. 891-897 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Miki,H.: "Association of ash/grb2 with dynamin through the src homology 3 domain." The Journal of Biological Chemistry.269. 5489-5492 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Terasawa,H.: "Solution Structure of human insulin-like growth factor II;recognition sites for receptors and binding proteins." The EMBO Journal. 13. 5590-5597 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Ichikawa,S.: "Solution Structure of Cystein-Rich Domain of Protein Kinase Cα." Journal of Biochemistry. (in press).

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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