Methylarginines and Vascular Injury

这项提案的长期目标是建立蛋白质精氨酸甲基转移酶和 二甲基精氨酸二甲氨基水解酶(DDAH)，负责甲基精氨酸的酶 合成和代谢，作为内皮功能的关键调节因子。我们的假设是 心血管疾病患者血浆ADMA升高是DDAH活性的生物标志物 ADMA对血管内皮细胞的许多影响是直接通过 改变了DDAH-PRMT活性。我们已经证明，除了ADMA对 ENOS活性，DDAH通过ADMA非依赖性调节内皮NO的产生 涉及蛋白质甲基化和氨基酸代谢改变的机制。世界银行的目标是 目前的建议是：1.)明确ADMA在内皮细胞中的代谢途径； 确定DDAH调节内皮细胞蛋白-精氨酸的机制 甲基化，明确蛋白质甲基化对内皮细胞功能的影响；确定 脱氢表雄酮调节血管内皮细胞L-精氨酸代谢的机制 对血管内皮细胞产生一氧化氮的影响；确定通过哪些机制 PRMT-DDAH-ADMA轴调节内皮功能和动脉粥样硬化敏感性。对于每一个 这些目标，结合了细胞、分子、生物物理和生理方法将 用体内外研究DDAH对血管内皮细胞功能的影响 模特们。这些研究的结果将提供基本的机理信息 PRMT-DDAH-ADMA轴调节细胞功能和 可能会带来治疗血管疾病的新方法。

The long term objective of this proposal is to establish Protein Arginine Methyltransferases and Dimethyarginine Dimethylaminohydrolase (DDAH), the enzymes responsible for methylarginine synthesis and metabolism, as a key regulator of endothelial function. It is our hypothesis that the increased plasma ADMA observed in cardiovascular disease is a biomarker of DDAH activity and that many of the endothelial affects attributed to ADMA are directly manifested through altered DDAH-PRMT activity. We have shown that in addition to the direct effects of ADMA on eNOS activity, DDAH modulates endothelial NO production through ADMA independent mechanisms involving altered protein-methylation and amino acid metabolism. The goals of the current proposal are to: 1.) identify the pathways of ADMA metabolism in the endothelium; 2.) determine the mechanisms through which DDAH modulates endothelial protein-arginine methylation and define the effects of protein methylation on endothelial function; 3.) determine the mechanisms through which DDAH regulates endothelial L-arginine metabolism and the consequences on endothelial NO production; and 4.) identify the mechanisms through which the PRMT-DDAH-ADMA axis regulates endothelial function and atherosusceptibility. For each of these aims, a combination of cellular, molecular, biophysical and physiological approaches will be used to characterize the effects of DDAH on endothelial function using in vitro and in vivo models. Results from these studies will provide fundamental mechanistic information regarding the mechanisms through which the PRMT-DDAH-ADMA axis modulates cellular function and may lead to new approaches to treat vascular disease.