| Project/Area Number |
06276105
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Research Institution | Tokyo Metropolitan Organization for Medical Research |
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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)
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| 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)
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| Project Period (FY) |
1994 – 1997
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| Project Status |
Completed (Fiscal Year 1998)
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| 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)
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| Keywords | three 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 |
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| 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.
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