Beta arrestin and NO signaling in endothelial cells

内皮细胞中的β抑制蛋白和NO信号传导

• 批准号: 7714554
• 负责人: Neetu Sud
• 金额: $ 8.48万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7714554
• 关键词: ARRB2; Adaptor Signaling Protein; Arginine; Arrestin Beta 1; Atherosclerosis; Attenuated; Binding; Blood Vessels; Blood flow; CREB1 gene; Calcineurin; Calcium; Cardiopulmonary; Cardiovascular Diseases; Cell membrane; Complex; Consensus; Coupled; Cyclic AMP; Cytosol; Data; Diabetes Mellitus; Disease; Doctor of Philosophy; Elements; Endothelial Cells; Enzymes; Event; Figs - dietary; Functional disorder; G-Protein-Coupled Receptors; G-substrate; Gene Expression; Generations; Genetic Transcription; HIV; Heat-Shock Proteins 90; Homeostasis; Human; Hypertension; In Vitro; Injury; Instruction; Location; Lung; Mediating; Mediator of activation protein; Membrane; Metabolism; Modeling; Molecular; Mutate; Nitric Oxide; Nitric Oxide Synthase; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Phosphorylation; Physiology; Plasma; Play; Production; Protein Dephosphorylation; Proteins; Pulmonary Hypertension; Reaction; Recombinants; Regulation; Relative (related person); Research; Role; SRC gene; Secondary to; Shunt Device; Signal Transduction; Small Interfering RNA; Superoxides; Testing; Tissues; Tyrosine; adenoviral-mediated; base; beta-arrestin; copper zinc superoxide dismutase; hemodynamics; human NOS3 protein; in vivo; inhibitor/antagonist; overexpression; prevent; response; uptake

DESCRIPTION (provided by applicant): An important factor in regulating eNOS activity is its localization. It has been demonstrated that the plasma membrane is the most efficient location for eNOS to produce NO in endothelial cells. Further, many studies have shown that HSP90 plays an important role in increasing eNOS mediated NO generation. However the molecular mechanisms by which eNOS-HSP90 interactions are stimulated are not fully understood. Beta arrestins are ubiquitous multifunctional adaptor proteins of G-protein coupled receptors (GPCR). Beta arrestin 2 has recently been shown to bind to HSP90. However, the role of beta arrestins in regulating HSP90-eNOS interactions has not been resolved. Our preliminary data indicate that a ternary complex of beta arrestin 1 or 2, HSP90 and eNOS is formed by calcium activating agents and is attenuated by calcium inhibiting agents. Overexpression of either beta arrestin or beta arrestin 2 results in higher NO production and lower eNOS-derived superoxide generation while the double knockdown of beta arrestin 1&2 expression attenuates NO production and augments eNOS-derived superoxide generation. These data have led us to form the hypothesis that beta arrestin promotes eNOS-HSP90 complex formation in the membrane by mediating the membrane translocation of HSP90. Oxidative stress mediates the pathogenesis of many cardiovascular diseases. Our preliminary data indicate that oxidative stress decreases the expression of beta arrestin. Its expression is decreased in our shunt model of pulmonary hypertension. We have previously shown that in these lambs, eNOS is uncoupled as evidenced by eNOS HSPO interactions are reduced, superoxide generation is enhanced. We hypothesize that a decrease in beta arrestin expression contributes to eNOS uncoupling in pulmonary hypertension. It has been shown that overexpression of CuZnSOD attenuates endothelial injury but the exact mechanisms of such protection remain unclear. Our preliminary data show that overexpression of SODl increases beta arrestini expression, enhances eNOS-HSP90 interactions and increases NO production and beta arrestini expression is increased in lambs treated with SODL We hypothesize that eNOS uncoupling can be restored by SODl overexpression via beta arrestin. RELEVANCE (See instructions); Our research is aimed at identifying the signaling mechanism of regulation of the enzyme eNOS, which produces nitric oxide, the molecule of fundamental importance for vascular homeostasis. In particular we trying to identify the cellular factor responsible for maintaining eNOS in the optimal "coupled" state when It generates NO and not superoxide, which is a key mediator of endothelial dysfunction associated with diseases like hypertension. diabetes, atherosclerosis etc.

描述（由申请人提供）：调节eNOS活性的一个重要因素是其定位。已经证明，质膜是eNOS在内皮细胞中产生NO的最有效位置。此外，许多研究表明，HSP 90在增加eNOS介导的NO产生中起重要作用。然而，刺激eNOS-HSP 90相互作用的分子机制尚未完全了解。β抑制蛋白是G蛋白偶联受体（GPCR）的普遍存在的多功能衔接蛋白。β抑制蛋白2最近被证明与HSP 90结合。然而，β抑制蛋白在调节HSP 90-eNOS相互作用中的作用尚未得到解决。我们的初步数据表明，β抑制蛋白1或2，HSP 90和eNOS的三元复合物是由钙激活剂形成的，并被钙抑制剂减弱。β抑制蛋白或β抑制蛋白2的过表达导致更高的NO产生和更低的eNOS衍生的超氧化物产生，而β抑制蛋白1和2表达的双重敲低减弱NO产生并增加eNOS衍生的超氧化物产生。这些数据使我们形成了这样的假设，即β抑制蛋白通过介导HSP 90的膜转位促进eNOS-HSP 90复合物在膜中的形成。氧化应激介导许多心血管疾病的发病机制。 我们的初步数据表明，氧化应激降低β抑制蛋白的表达。其表达在我们的肺动脉高压分流模型中减少。我们以前已经表明，在这些羔羊中，eNOS是解偶联的，如eNOS HSPO相互作用减少所证明的，超氧化物生成增强。我们假设β抑制蛋白表达的减少有助于肺动脉高压中eNOS的解偶联。研究表明，CuZnSOD的过度表达可减轻内皮损伤，但这种保护作用的确切机制尚不清楚。我们的初步数据显示，SOD 1的过表达增加β抑制蛋白表达，增强eNOS-HSP 90相互作用并增加NO产生，并且在用SODL处理的羔羊中β抑制蛋白表达增加。我们假设，eNOS解偶联可以通过SOD 1过表达经由β抑制蛋白恢复。相关性（参见说明）;我们的研究旨在确定eNOS酶调节的信号机制，eNOS酶产生一氧化氮，一氧化氮是血管稳态的重要分子。特别是，我们试图确定负责维持eNOS在最佳的“耦合”状态时，它产生NO，而不是超氧化物，这是一个关键的调解人的内皮功能障碍与疾病，如高血压的细胞因子。糖尿病、动脉粥样硬化等。