| Project/Area Number |
14208076
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Hokkaido University |
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Principal Investigator |
INAGAKI Fuyuhiko Hokkaido University, Grad.School of Pharm.Sci., Professor, 大学院・薬学研究科, 教授 (70011757)
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| Co-Investigator(Kenkyū-buntansha) |
OGURA Kenji Hokkaido University, Grad.School of Pharm.Sci., Instructor, 大学院・薬学研究科, 助手 (50270682)
HORIUCHI Masataka Hokkaido University, Grad.School of Pharm.Sci., Instructor, 大学院・薬学研究科, 助手 (90322825)
小椋 賢治 北海道大学, 大学院・薬学研究科, 助手 (14780469)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥51,350,000 (Direct Cost: ¥39,500,000、Indirect Cost: ¥11,850,000)
Fiscal Year 2004: ¥16,510,000 (Direct Cost: ¥12,700,000、Indirect Cost: ¥3,810,000)
Fiscal Year 2003: ¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000)
Fiscal Year 2002: ¥16,770,000 (Direct Cost: ¥12,900,000、Indirect Cost: ¥3,870,000)
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| Keywords | phagocyte oxidation system / protein-protein interaction / SH3 domain / activation mechanism / auto inhibition / p47 tandem SH3 domains / PRR peptide / 新規NMR技術 |
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| Research Abstract |
The phagocyte NADPH oxidase, a defense system for microbial infection comprises membrane bound flavocytochrome cyt b_<558> (gp91^<phox> and p22^<phox>) and cytosolic factors including p67^<phox>, p47^<phox>, p40^<phox> and small GTPase Rac. Upon microbial invasion, the cytosolic factors are translocated to the membrane to bind to cyt b_<558>, which activates a redox-core of cyt b_<558> and generates O_2-, a precursor of microbicidal oxidants. The activation of the NADPH oxidase is tightly regulated through conformation change of the tandem SH3 domain of p47^<phox>.
Recently, the crystal structure of the tandem SH3 domains in the autoinhibited form (Groemping et al., Cell,2003, Yuzawa et al., Genes Cells,2004, J Biol Chem.,2004) and the activated form (Groemping et al., Cell,2003) were determined. Interestingly, the tandem SH3 domains formed an intertwisted dimer in the crystal state but the splitted half was considered to be physiologically relevent. Here, we report the three dimensional structures of the masked and unmasked states of the tandem SH3 domains by NMR. Furthermore, we elucidated the global conformation change of the tandem SH3 domains from the masked to unmasked states by small angle X-ray scattering. Based on the present structural data, we discussed the activation mechanism of the phagocyte NADPH oxidase.
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