CaMK II Variants and Vascular Smooth Muscle Function

描述（由申请人提供）：该项目的总体目标是确定单个CaM激酶II变异在平滑肌调节中的功能，以全面了解该蛋白激酶家族在平滑肌中的功能。我已经从雪貂主动脉cDNA文库中鉴定出CaM激酶II γ的六种变体作为cDNA。它们的体外功能在自磷酸化和底物磷酸化方面有所不同。这表明它们在细胞中的功能也会有所不同。拟议的研究将以特定的目的进行：i)验证从主动脉cDNA文库中鉴定出的CaM激酶II γ的所有六种变体在平滑肌组织中以蛋白质形式表达的假设；2)验证CaM激酶II γ的不同变体具有不同功能的假设；3)验证CaM激酶II δ的多种变体也表达并参与平滑肌调节的假设。使用的技术包括克隆、定量RT-PCR、质谱测序、反义敲除、共聚焦显微镜和蛋白激酶测定。

DESCRIPTION (provided by applicant): The general aim of the project is to determine the function of individual CaM kinase II variants in smooth muscle regulation toward the goal of obtaining a complete understanding of the function of this family of protein kinases in smooth muscle. I have already identified six variants of CaM kinase II gamma, as cDNAs, from a ferret aorta cDNA library. Their in vitro functions differ in terms of autophosphorylation and substrate phosphorylation. This suggests that they will also function differentially in the cell. The proposed study will be undertaken with the specific aims i) To test the hypothesis that all six variants of CaM kinase II gamma identified as cDNAs from an aorta cDNA library are expressed as protein in smooth muscle tissue; 2) To test the hypothesis that different variants of CaM kinase II gamma have differential functions; 3) To test the hypothesis that multiple variants of CaM kinase II delta are also expressed and contribute to the regulation of smooth muscle. Technologies to be used include, cloning, quantitative RT-PCR, mass spec sequencing, antisense knockdown, confocal microscopy and protein kinase assays.