Studies on the effect of caveolin-analogues on the structure and function of caveolae

内皮源性一氧化氮（NO）是心血管系统多种功能的重要介质。在过去的几年里，一个概念已经发展起来，即eNOS的激活/失活可以通过磷酸化和与几种蛋白质（包括Cav-1）的区域特异性蛋白质-蛋白质相互作用来调节。在初步实验中，通过诱变小窝蛋白支架结构域（CSD），从抑制功能的角度分析了eNOS与Cav-1之间的相互作用，并开发了一种新的细胞渗透性eNOS激活肽（Cav-1- f92a）。因此，我们提出了一种假设，即不抑制eNOS的Cav-1突变体或细胞渗透性eNOS激活剂会拮抗Cav-1对eNOS的抑制作用，促进NO释放，改善内皮细胞健康和血液流动。为了更详细地研究这一重要相互作用的调节，提出了以下具体目标：将Cav-1-F92A重组为野生型和敲除cav -1的内皮细胞和血管；2)在内皮细胞中诱导表达Cav-1-F92A的转基因小鼠。总的来说，这项工作将有助于理解EC中eNOS调控所需的分子机制。科学意义：这项研究与公众健康相关，因为内皮功能障碍是大多数心血管疾病的常见表现。这项拨款支持的研究可能有助于发现减少心脏病和改善心血管疾病患者生活质量的新药。

Endothelium-derived nitric oxide (NO) is a critical mediator of diverse functions in the cardiovascular system. During the last years a concept had developed that the eNOS activation/inactivation can be regulated by phosphorylation and locale specific protein-protein interactions with several proteins including caveolin-1 (Cav-1). In preliminary experiments, the interaction between eNOS and Cav-1 has been dissected from its inhibitory function using mutagenesis of the caveolin scaffolding domain (CSD) and a novel cell permeable, eNOS activator peptide was developed (Cav-1-F92A). Thus, the hypothesis arises that a mutant of Cav-1 that does not inhibit eNOS or a cell permeable eNOS activator will antagonize the inhibitory role of Cav-1 on eNOS and promote NO release and improve endothelial cell health and blood flow. To examine the regulation of this important interaction in more detail, the following specific aims are proposed: 1.) Reconstitute Cav-1-F92A into wild-type and Cav-1-knockout endothelial cells and vessels and 2.) generate transgenic mice that inducibly express Cav-1-F92A in the endothelium. Collectively, this work will facilitate the understanding of the molecular machinery required for eNOS regulation in EC. Scientific Relevance: This research is relevant to public health since endothelial dysfunction is a common manifestation of most cardiovascular diseases. Research supported by this grant may help identify new drugs that reduce heart disease and improve the quality of life of people suffering with cardiovascular disease.