Regulation of Myosin Light Chain Kinase by Phosphorylat*

描述（由申请人提供） 肌球蛋白轻链激酶（MLCK）是调节平滑肌细胞增殖的关键元件。 肌肉收缩。特别是MLCK的磷酸化，已经成为一个重要的研究领域。 可能将收缩性与其他信号通路联系起来的事件。在这 应用程序，我们的目标是填补我们的知识的差距，如何网站的具体 磷酸化影响MLCK活性。我们的长期目标是了解 平滑肌细胞内的信号转导过程及其机制 其规定。具体而言，我们建议（i）开发磷酸化特异性 抗体，以监测细胞中MLCK的磷酸化水平， MLCK磷酸化与心肌收缩特性的关系 （ii）为了检验平滑肌细胞/非肌肉细胞的磷酸化 MLCK通过MAPK影响酶的活性并探讨其作用机制 研究MLCK磷酸化对其构象的影响;以及（iii） 确定MAPK和PKA的磷酸化如何影响MLCK与 平滑肌和非肌肉收缩机制的组成部分， 原位定位。拟议的研究是一个合乎逻辑的延伸， 由NIH资助的正在进行的计划项目赠款（P01-AR41637），标题为 "平滑肌调节的分子机制。"拟议的合作 不仅可以提高申请人的研究计划（C. - L.阿尔伯特·王， 博士，美国首席研究员），但也将有利于科学 外国合作者的利益，亚历山大沃罗特尼科夫博士，并强烈 影响了国外实验室的研究。本报告中描述的工作 提案将提供信息，以扩大正在进行的 研究MLCK的磷酸化调控，为MLCK的研究提供了更好的理论基础。 了解平滑肌调节。

DESCRIPTION (provided by applicant) Myosin light chain kinase (MLCK) is a key element in the regulation of smooth muscle contraction. Phosphorylation of MLCK, in particular, has emerged as an event that may link contractility to other signaling pathways. In this application we aim to fill the gaps in our knowledge of how site-specific phosphorylation affects the MLCK activity. Our long term goal is to understand the signal transduction process in smooth muscle cells and the mechanisms for its regulation. Specifically, we propose (i) to develop phospho-specific antibodies to monitor the phosphorylation level of MLCK in cells and to assess the relationship between phosphorylation of MLCK and the contractile properties of smooth/non-muscle cells; (ii) to test the hypothesis that phosphorylation of MLCK by MAPK affects the activity of the enzyme and to explore the mechanism by investigating effects of MLCK phosphorylation on its conformation; and (iii) to determine how phosphorylation by MAPK and PKA affects interaction of MLCK with components of smooth muscle and non-muscle contractile machinery by elucidating the in situ localization. The proposed research is a logical extension of the ongoing Program Project Grant funded by NIH (P01-AR41637) under the title of "Molecular Mechanisms of Smooth Muscle Regulation." The proposed collaboration will not only enhance the research program of the applicant (C.-L. Albert Wang, Ph.D., the US Principal Investigator), but will also benefit the scientific interests of the Foreign Collaborator, Dr. Alexander Vorotnikov, and intensely impact the research conducted in the Foreign Laboratory. Work described in this proposal will provide information to extend the applicability of the ongoing studies to the regulation of MLCK by phosphorylation, and afford a better understanding of smooth muscle regulation.