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变体在平滑肌调节中的功能，以达到全面了解这一蛋白激酶家族在平滑肌中的功能的目标。我已经从雪貂主动脉的cDNA文库中鉴定出了六种CaM激酶II伽马的变异体，即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是不同的。这项研究的具体目的是：1)检验从主动脉cDNA文库中鉴定为cDNA的所有6个CaM KK IIγ变异体均以蛋白质形式在平滑肌组织中表达的假设；2)检验不同Cam Kinase II GAMA变异体具有不同功能的假说；3)检验CaM K II Delta的多种变异体也表达并参与平滑肌调节的假说。将使用的技术包括克隆、定量RT-PCR、质谱仪测序、反义基因敲除、共聚焦显微镜和蛋白激酶分析。

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.