Phosphoproteome and Ang II-induced VSMC Gene Expression

血管平滑肌细胞（VSMC）表型在健康和 疾病状态。然而，VSMC表型调节的分子机制尚不清楚。 在血管紧张素II刺激的VSMC模型系统中，我们发现磷酸化蛋白质组和 转录组细胞骨架蛋白|3-肌动蛋白被磷酸化，导致假设， 磷酸化可以改变肌动蛋白动力学，这是细胞运动、生长和基因表达所必需的过程 变化基因表达谱分析表明，几种细胞表面受体，转录调节因子和 参与器官损伤的蛋白质是差异表达的。启动子的生物信息学分析 这些基因的区域显示出遗传上保守的DMAc/s元件。我们推测这些 基因组调控元件与转录调节因子结合，转录调节因子是血管紧张素II诱导信号的靶点 转导和改变基因表达。 本研究的短期目标是：（i）鉴定血管紧张素II处理的VSMCs中磷酸化的蛋白靶点 通过免疫纯化和质谱（MS）。我们将测试磷酸化的网络 通过基因表达变化调节VSMC表型的蛋白质，（ii）确定Ang II的位点， 诱导的磷酸化和磷酸化β-肌动蛋白影响VSMC增殖的机制， （iii）确定基因的Ang II-反应性的分子基础。 我们的长期目标是了解血管紧张素II（Ang II）对VSMC功能的调节， VSMC表型调控的分子机制。人类主要疾病的发病机制， 动脉粥样硬化和血管成形术后再狭窄涉及VSMC表型转换。

Vascular smooth muscle cell (VSMC) phenotypes display remarkable heterogeneity in health and disease states. Yet the molecular mechanisms underlying modulation of VSMC phenotypes are not known. In Ang ll-stimulated VSMC model system, we find global changes in both the phosphoproteome and transcriptome. The cytoskeletal protein |3-actin is phosphorylated, leading to the hypothesis that phosphorylation could alter actin-dynamics, a process required for cell motility, growth and gene expression changes. Gene expression profiling indicated that several cell surface receptors, transcription regulators and proteins involved in organ damage are differentially expressed. Bioinformatics analysis of the promoter regions of these genes showed phylogenetically conserved DMAc/s-elements. We speculate that these genome regulatory elements bind transcriptional regulators, which are targets of Ang ll-induced signal transduction and alter gene expression. The short-term goals of this study are: (i) Identify phosphorylated protein targets in Ang ll-treated VSMCs by using immuno-purification and mass spectrometry (MS). We will test the network of phosphorylated proteins that modulate VSMC phenotype through gene expression changes, (ii) Determine site(s) of Ang ll- induced phosphorylation and the mechanism by which phosphorylation of p-actin affects VSMC proliferation, migration and gene expression, (iii) Define the molecular basis for Ang ll-responsiveness of genes. Our long-term goal is to understand regulation of VSMC function by angiotensin II (Ang II) and the molecular mechanism of phenotypic modulation of VSMC. Pathogenesis of major human diseases such as atherosclerosis and post-angioplasty restenosis involves VSMC phenotype switching.