Structural basis for auto-inhibition mechanism of Rho-kinase
| Project/Area Number |
13680742
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biophysics
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| Research Institution | Yokohama City University |
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Principal Investigator |
SHIMIZU Toshiyuki Science of Biological Supramolecular Systems Assoc. Prof., 総合理学研究科, 助教授 (30273858)
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| Co-Investigator(Kenkyū-buntansha) |
HAKOSHIMA Toshio Nara Inst. Of Sci. and Tech. Structural Biology Lab. Prof., 情報科学研究科, 教授 (00164773)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Rho-kinase / Rho-signalling / auto-inhibition / X-ray analysis / tertiary structure / ERM protein / Merlin / FERM domain / Rho-キナーゼ / コイルドコイル / DHドメイン / Rho |
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| Research Abstract |
Rho-kinase containing Kinase domain and part of coiled-coil domain is expressed by baculovirus systems as a fusion protein with glutathione-S-transferase. The protein is purified by several column chromatography steps and was finally obtained as a single band, stained with Coomassie brilliant blue, in SDS polyacrylamide gel electrophoresis. We collected X-ray small angle scattering data to check the protein. The data show the protein solution is mono-disperse and protein forms tetramer in solution. All the crystallization experiments were carried out with the hanging-drop vapor-diffusion method using 24-well tissue-culture plates (sumitomo Bakelite Co.). We searched various crystallization conditions extensively, but failed to get crystals until now. ERM (ezrin/radixin/moesin) proteins recognize the cytoplasmic domain of adhesion molecules in the formation of the membrane-associated cytoskeleton. We report the crystal structure of the radixin FERM domain completed with the ICAM-2 cytop
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lasmic peptide. Mutations analyses based on the crystal structure reveal the determinant elements of recognition and provide the first insight into the physical link between adhesion molecules and ERM proteins.
Neurofibromatosis type 2 (NF2) is a dominantly inherited disease associated with the central nervous system. The NF2 gene product merlin is a tumor suppressor and its mutation or inactivation causes this disease.
We report here the crystal structure of the merlin FERM domain containing a 22-residue a helical segment. The structure reveals that the merlin FERM domain consists of three subdomains displaying notable features of the electrostatic surface potentials, although the overall surface potentials similar to those of ERM proteins indicate its electrostatic membrane association. The structure also suggests the inactivation mechanisms caused by the pathogenic mutations associated with NF2.
Moreover, we get the crystals of PhoGDI complied with ERM protein. Structure determination is in progress. Less
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Report
(3 results)
Research Products
(19 results)
