| Project/Area Number |
05453210
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Structural biochemistry
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| Research Institution | Tokyo Metropolitan Institute of Medical Science |
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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) |
OGURA Kenji Tokyo Metropolitan Institute of Medical Science Researcher, 生理活性物質研究部門, 研究員 (50270682)
HATANAKA Hideki Tokyo Metropolitan Institute of Medical Science Researcher, 生理活性物質研究部門, 研究員 (00260331)
KOHDA Daisuke Tokyo Metropolitan Institute of Medical Science Researcher, 生理活性物質研究部門, 研究員 (80186618)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1994: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1993: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Keywords | NMR / distance geometry / solution structure / erabutoxin b / heregulin alpha / IGF-II / SH3 domain / proline-rich peptide |
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| Research Abstract |
We constructed a computer program system to determine the high reolution solution structure of proteins by NMR.The program were applied to determine the structure of heregulin, which is a ligand to ErbB4 receptor which is similar to EGF receptor but is important for regemeration of central nervous system and tumor fotmation. The structure is quite similar to EGF and we expect that heregulin also has similar binding surface to EGF.The patch on the putative receptor binding surface is quite different from that of EGF,supporting that these ligand bind exclusively to cognate receptors. We also determind the solution structure of IGF-II.IGF-II is similar to insulin and IGF-I.We found that the binding sites for insulin/IGF-I receptor and IGF-II receptor are located on the opposite surface of IGF-II.We extended our structural study to the protein interaction in the cytosolic state. SH2 and SH3 are structural module integrated in the signal transduction system. SH2 binds to the activated receptors by the binding to a phosphotyrosine residue in the receptor. Src-homology 3 (SH3) domains mediate signal transduction by binding to proline-rich motifs in target proteins involved in signal transduction system. We have determined the high-resolution structure of the SH3 domains of PLCgamma and Grb2. The three dimensional structure of SH3 domains are quite similar in spite of the low sequence homology (20%). But, owing to low affinity of proline-rich peptide to those SH3 domains, we could not determind the complex structures. Fortunately, VPPPVPPRRR peptide derived from Sos bound to Grb2 N-SH3 with high affinity and we have determined the high-resolution NMR structure of the complex. The NMR data show that the peptide adopts the conformation of a left-handed polyproline type II helix and interacts with three major sites on the SH3 domain. The orientation of the bound peptide is opposite to that of proline-rich peptides bound to the SH3 domains of Abl, Fyn and p85.
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