Regulation of Myosin Light Chain Kinase by Phosphorylat*

磷酸化物对肌球蛋白轻链激酶的调节*

基本信息

批准号：
6697070
负责人：
CHIH-LUEH Albert WANG
金额：
$ 3.2万
依托单位：
BOSTON BIOMEDICAL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-01-01 至 2005-12-31
项目状态：
已结题

项目摘要

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.

描述（由申请人提供）肌球蛋白轻链激酶（MLCK）是平滑肌细胞调节的关键元件。肌肉收缩。特别是 MLCK 的磷酸化已成为一种可能将收缩性与其他信号通路联系起来的事件。在这个我们的应用程序旨在填补我们在如何针对特定地点进行操作方面的知识空白磷酸化影响 MLCK 活性。我们的长期目标是了解平滑肌细胞的信号转导过程及其机制其监管。具体来说，我们建议 (i) 开发磷酸盐特异性抗体监测细胞中 MLCK 的磷酸化水平并评估 MLCK磷酸化与收缩特性的关系平滑/非肌肉细胞； (ii) 检验磷酸化的假设 MLCK通过MAPK影响酶的活性并探讨其机制研究 MLCK 磷酸化对其构象的影响； (iii) 至确定 MAPK 和 PKA 的磷酸化如何影响 MLCK 与通过阐明平滑肌和非肌肉收缩机制的组成部分原位定位。所提出的研究是该研究的逻辑延伸由 NIH (P01-AR41637) 资助的正在进行的计划项目拨款，标题为 “平滑肌调节的分子机制。”拟议的合作不仅会增强申请人的研究计划（C.-L. Albert Wang，博士，美国首席研究员），但也将有利于科学外国合作者亚历山大·沃罗特尼科夫博士的兴趣，并强烈影响外国实验室进行的研究。本文描述的工作提案将提供信息以扩展正在进行的适用性研究磷酸化对 MLCK 的调节作用，并提供更好的了解平滑肌调节。