| Project/Area Number |
61480112
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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 |
Neurophysiology and muscle physiology
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| Research Institution | National Institute for Physiological Sciences |
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Principal Investigator |
EBASHI Setsuro Director General of Physiological Sciences, 生理学研究所, 所長 (10009863)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIZAKI Yasuki Postdoctral Fellow of JSPS, 生理学研究所, 学術振興会特別研究員 (90183003)
SUZUKI Masashi Research Associate, 生理学研究所, 助手 (70187764)
MIKAWA Takashi Research Associate, 生理学研究所, 助手 (50143417)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1987: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1986: ¥4,500,000 (Direct Cost: ¥4,500,000)
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| Keywords | Regulation in smooth muscle / Leiotonin / ミオシン軽鎖キナーゼ |
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| Research Abstract |
A new method of preparing leiotonin was developed. It was incidentally found that dimethyl sulfoxide (DMSO) is a very useful agent for protein fractionation, not only for the above purpose but also for general ones. Using DMSO, a fraction exhibiting much larger actomyosin-activating activity, or leiotonin activity, than myosin light chain kinase (MLCK) activity (L/K > 100, taking L/K of 0.1 M extract as 1) was obtained both from chicken gizzard and boving stomach without particular treatment. Using special procedures, a preparation void of MLCK avtivity (L/K > 3,000) could be obtained. The component responsibe for the leiotonin activity was a protein of 155 kDa, indistinguishable from MLCK in its molecular weight.
A fragment produced by trypsin under specified conditions exhibited also a definite leiotonin activity without MLCK activity. This may correspond to the old preparation of leiotonin.
The primary sequence of the 155 kDa component responsible for leiotonin activity (and/or MLCK activity) has been determined up to 120 kDa from the C- terminal.
As a whole the idea that smooth muscle regulation is mediated by leiotonin has further been substantiated.
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