| Project/Area Number |
03454541
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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 |
物質生物化学
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| Research Institution | The Tokyo Metropolitan Institute of Medical Science |
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Principal Investigator |
YAHARA Ichiro Tokyo Met. Inst. Med. Sci., Vice-Dir., 副所長 (60109957)
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| Co-Investigator(Kenkyū-buntansha) |
MIYATA Yoshihiko TMIMS.Dept. Cell. Biol., Res. Sci., 細胞生物学研究部門, 研究員 (70209914)
IIDA Kazuko TMIMS.Dept. Cell. Biol., Res. Sci., 細胞生物学研究部門, 研究員 (40151229)
MATSUMOTO Seiji TMIMS.Dept. Cell. Biol., Res. Sci., 細胞生物学研究部門, 研究員 (40190532)
MINAMI Yasufumi TMIMS.Dept. Cell. Biol., Res. Sci., 細胞生物学研究部門, 研究員 (40181953)
小安 重夫 東京都臨床医学総合研究所, 細胞生物, 研究員 (90153684)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1992: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1991: ¥4,400,000 (Direct Cost: ¥4,400,000)
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| Keywords | stress proteins / HSP90 / calmodulin-binding domain / dimerization domain / molecular chaperone / casein kinase II / budding yeast / temperature-sensitive mutants / カルパイン処理 / 酵素の失活 / 会合体形成 / アクチン結合タンパク質 / カルモデュリン / 酵母温度感受性変異株 / 合成致死変異 / グルココルチコイド受容体 / カゼインキナ-ゼII / 複合体形成 |
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| Research Abstract |
1) Domain structure of HSP90 : The calmodulin-binding domain of HSP90 was identified by cross-linking of HSP90 and Ca-calmodulin with a chemical cross-linker, EDC, followed by CNBr treatment. The domain consists of 21 amino acid residues which fold into an amphiphilic alpha-helix, This identification was confirmed by chemically syntheizing the paptide and examining its binding ability to calmodulin. Next, proteolytic cleavage of C-terminus of HSP90 was expressed in reticulocyte lysate, it formed dimer. When a mutated HSP90 lacking of its C-terminal 49 amino acids was expressed, no dimer was formed. These results suggest that the calboxy-terminal region is necessary for its dimerization.
2) We found that HSP90 protects casein kinase II from self-aggregation and inactivation.
3) We isolated five HSP90 ts-mutants of the budding yeast, Saccharomyces cerevisiae. Phenotypes expressed at the non-permissive temperature were extensively analyzed.
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