| Project/Area Number |
16209007
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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 |
General pharmacology
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| Research Institution | Gunma University |
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Principal Investigator |
KOHAMA Kazuhiro Gunma University, Department of Molecular and Cellular Pharmacology, Professor (30101116)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIKAWA Ryoki Gunma University, Department of Molecular and Cellular Pharmacology, Assoc. Prof. (20212863)
NAKAMURA Akio Gunma University, Department of Molecular and Cellular Pharmacology, Assoc. Prof. (30282388)
YOSHIYAMA Shinji Gunma University, Department of Molecular and Cellular Pharmacology, Assist Prof. (30396650)
TANAKA Hideyuki Gunma University, Department of Faculty of Medicine, Research Associate (70343085)
NASU Ritsuko Takasaki University, Department of Health & Welfare Nutritia, Assist Prof. (60383147)
登坂 雅彦 群馬大学, 大学院・医学系研究科, 助手 (40323357)
熊谷 啓之 群馬大学, 医学系研究科, 助手 (20321945)
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| Project Period (FY) |
2004 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥48,750,000 (Direct Cost: ¥37,500,000、Indirect Cost: ¥11,250,000)
Fiscal Year 2007: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2006: ¥12,740,000 (Direct Cost: ¥9,800,000、Indirect Cost: ¥2,940,000)
Fiscal Year 2005: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2004: ¥13,520,000 (Direct Cost: ¥10,400,000、Indirect Cost: ¥3,120,000)
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| Keywords | Actin / Myosin / smooth muscle / Regulation / migration / 薬理学 / 蛋白質 / 循環器・高血圧 / リン酸化 / ブレビスタチン / 血小板成長因子 |
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| Research Abstract |
Myosin light-chain kinase (MLCK) of smooth muscle consists of an actin-binding domain at the N-terminal the catalytic domain in the central portion, and the myosin-binding domain at the C-terminal. The kinase activity is mediated by the catalytic domain that phosphorylates the myosin light-chain of 20 kDa (MLC20), activating smooth muscle myosin to interact with actin. Although the regulatory role of the kinase activity is well established, the role of non-kinase activity derived from actin-binding and myosin-binding domains remains unknown. We hypothesized that the actin-myosin interaction of smooth muscle could be activated by the non-kinase activity independently of MLC20 phosphorylation. We expressed cDNA of MLCK as a recombinant protein, and mutated it to lose the kinase activity, while keeping non-kinase activity unaffected. With the mutant, we detected an activating effect on the actin-myosin interaction. We also down-regulated MLCK expression by its RNAi in the cultured smooth muscle cells, to observe the lack of the activation. The rescue experiments with the MLCK-deficient cells demonstrated that MLC20 phosphorylation is not obligatory for the smooth muscle to contract.
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