| Project/Area Number |
10680618
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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 |
Functional biochemistry
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| Research Institution | Sapporo Medical University |
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Principal Investigator |
YOSHINO Hidenori Sapporo Medical University, Medical School, Associate Prof., 医学部, 助教授 (70045524)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1998: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | calcineurin / calmodulin / calcium / X-ray solution scattering / UV-difference absorption spectrum / 紫外部差スペクトル / X線溶液散乱法 / ニッケル |
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| Research Abstract |
Solution structure of protein phosphatase calcinerin have been studied by using a solution X-ray scattering method. the radius of gyration (Rg) of calcineurin without metal ions or with MgィイD12+ィエD1, in which calcineurin is in the off-state, are the same values of 43.1A respectively. While, the Rg values of calcineurin at the on-state which are induced by CaィイD12+ィエD1 or CaィイD12+ィエD1- calmodulin have changed to 5704A and 64.3A, respectively. Analyses of pair distribution function for the scattering data indicated that calcineurin and MgィイD12+ィエD1-calcineurin have oval shape with molecular length in 93A, and that for CaィイD12+ィエD1-calmodulin-calcineurin complex approximates three-domain structure with length in 107A. Energy transfer from tryptophany1 residues in calcineurin to a dansy1 chloride attached covalently on calmodulin have occurred indicating the domain of A subunit is close to a domain induced by calmodulin binding. The Kratky plots of the scattering date from calcineurin suggest that tertiary structure of calcineurin at the off-state is similar to that of the on-state. Furthermore, UV absorption spectrum of calcineurin with CaィイD12+ィエD1 can superimpose on that obtained without CaィイD12+ィエD1, exhibiting little change in the tertiary structure between them.
The findings obtained here suggest that the on/off state of calcineurin should be regulated by adjustment of the relative position of domains without a change in the tertiary structure.
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