Regulatory Factor of Smooth Muscle Contraction: Relation of Leiotonin with Myosin Light Chain Kinase

平滑肌收缩的调节因素：降钙素与肌球蛋白轻链激酶的关系

• 批准号: 61480112
• 负责人: EBASHI Setsuro
• 金额: $ 4.16万
• 依托单位: National Institute for Physiological Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61480112/
• 关键词: Regulation in smooth muscle; Leiotonin; ミオシン軽鎖キナーゼ

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.

提出了一种新的合成列黄素的方法。偶然发现二甲亚砜（DMSO）是一种非常有用的蛋白质分离剂，不仅适用于上述目的，而且适用于一般目的。使用DMSO，在未经特殊处理的情况下，从鸡胗和鸡胃中均可获得比肌球蛋白轻链激酶（MLCK）活性大得多的肌动球蛋白活化活性（leiotonin activation activity，或leiotonin activity） （L/K为100，以0.1 M提取物的L/K为1）。通过特殊的工艺，可以获得MLCK活性（L/K > 3000）的制备空隙。负责leiotonin活性的成分是一种分子量为155 kDa的蛋白质，与MLCK在分子量上没有区别。胰蛋白酶在特定条件下产生的片段也显示出一定的卵黄素活性，但没有MLCK活性。这可能与旧的列素制剂相对应。155 kDa的组成部分负责leiotonin活性（和/或MLCK活性）的初级序列已经从C-末端确定到120 kDa。从整体上看，平滑肌调节是由络合素介导的这一观点得到了进一步的证实。

项目成果

Ebashi,S.: In Heart Function and Metabolism.(eds.by N.S.Dhalla,G.N.Pirtce,R.E.Beamish)Martinus Nijhoff Publishing. (1987)

Ebashi,S.：心脏功能和代谢。（由 N.S.Dhalla、G.N.Pirtce、R.E.Beamish 编辑）Martinus Nijhoff 出版。

Ebashi, S.: "Actomyosin ATPase and superprecipitation: Their modification by rutin." Acta Biochim. Biophys. Hung.22(2-3). 295-306 (1987)

Ebashi, S.：“肌动球蛋白 ATP 酶和超沉淀：芦丁对它们的修饰。”

Ebashi,S.;Ogawa,Y.: "In Handbook of Experimental Pharmacology(eds.by G.V.R.Born,A.Farah,H.Herken,A.D.Welch)Springer-Verlag Berlin Heidelerg New York Tokyo" Calcium in drug actions., (1987)

Ebashi,S.；Okawa,Y.：“实验药理学手册（G.V.R.Born、A.Farah、H.Herken、A.D.Welch 编）Springer-Verlag 柏林 海德堡 纽约 东京”药物作用中的钙。，（

Ebashi, S.;Ogawa, Y..;G. V. R. Born;A. Farah;H. Herken;A. D. Welch: In Handbook of Experimental Pharmacology Springer-Verlag Berlin Heidelberg New York Tokyo. 31-56 (1988)

Ebashi，S.；小川，Y..；G.

Ebashi, S.;eds. by Kleinkauf, von D8hren Janicke: The Roots of Modern Biochemistry Walter de Gruyter & Co., Berlin, New York. 737-746 (1988)

Ebashi，S.；编辑。