eNOS-Protein Interaction in the Penis

阴茎中的 eNOS-蛋白质相互作用

• 批准号: 7130615
• 负责人: BILJANA MUSICKI
• 金额: $ 24.54万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7130615
• 关键词: NAD(P)H dehydrogenase; antioxidants; apolipoproteins; biomimetics; cardiovascular pharmacology; caveolins; enzyme activity; free radical oxygen; heat shock proteins; high density lipoproteins; hypercholesterolemia; immunoprecipitation; impotence; laboratory mouse; nitric oxide; nitric oxide synthase; penis erection; peroxynitrites; phosphorylation; protease inhibitor; protein protein interaction; vascular endothelium

DESCRIPTION (provided by applicant): Endothelial nitric oxide synthase (eNOS) is an important mediator of penile erection. Appropriate subcellular trafficking and protein-protein interactions are necessary for proper eNOS function. Among the most important eNOS-protein interactions that may affect eNOS regulation in the vasculature including the penile vascular bed, are eNOS binding with caveolin-1 and heat shock protein (Hsp)90. While caveolin-1 facilitates eNOS trafficking to caveolae and proper organization of the eNOS signaling complex, it interferes with subsequent biological actions of eNOS. Hsp90 facilitates eNOS activation and keeps coupled eNOS activity. Currently very little is known about the role and regulation of eNOS-protein interactions in the regulation of the enzyme's activity and its functional relevance in the vasculature including the penile vascular bed. The overall goal of the proposal is to investigate the molecular mechanisms of eNOS regulation in the penis by eNOS-protein interactions, and its physiological significance in penile erection. The investigation will involve evaluation of eNOS interactions with caveolin-1 and HspQO in a mouse dyslipidemic model as a paradigm for this investigation. We will first evaluate whether eNOS interactions with caveolin-1 and HspQO are impaired in hypercholesterolemia-induced erectile dysfunction (ED) under flaccid or erect conditions, in parallel with a defect in eNOS function. Next, we will evaluate whether preserving eNOS-protein interactions preserves eNOS function and erectile function. This hypothesis will be tested in dyslipidemic mice treated with L-4F, an apolipoprotein-A-l mimetic, which positively influences eNOS function in part by maintaining an optimal eNOS-protein interaction. Third, we will evaluate whether increased superoxide production by NAD(P)H oxidase and uncoupled eNOS underlies impaired eNOS-protein interaction and ED in the hypercholestero- lemic mouse. This will be tested in hypercholesterolemic mice with and without apocynin-induced inhibition of NAD(P)H oxidase. Elucidation of the molecular mechanisms of eNOS regulation is crucial for a better understanding of NO-mediated responses in vascular tissues under physiologic and pathophysiologic circumstances. The proposed investigation is expected to elucidate a novel molecular mechanism of eNOS regulation in the penis and its physiological significance in erection physiology. At the same time, it may suggest a specific eNOS regulatory pathway as a possible new molecular target for treating ED.

描述（由申请人提供）：内皮一氧化氮合酶（ENOS）是阴茎勃起的重要介体。适当的亚细胞运输和蛋白质 - 蛋白质相互作用对于适当的eNOS功能是必需的。在包括阴茎血管床在内的eNOS调节的最重要的eNOS-蛋白质相互作用之一是与小窝蛋白-1和热休克蛋白（HSP）90结合的eNOS结合。 Caveolin-1促进eNos运输到小屋和eNOS信号复合体的适当组织，但它会干扰ENOS的随后生物学作用。 HSP90促进ENOS激活并保持耦合的eNOS活性。目前，关于eNOS-蛋白质相互作用在调节酶活性及其在包括阴茎血管床（阴茎血管床）中的功能相关性中的作用和调节知之甚少。该提案的总体目标是通过eNOS蛋白质相互作用研究阴茎中eNOS调节的分子机制，及其在阴茎勃起中的生理意义。该研究将涉及对小鼠血脂症模型中的eNOS相互作用与小鼠1和HSPQO的评估，作为这项研究的范例。我们将首先评估在弱胆固醇血症诱导的勃起功能障碍（ED）中，在易受耐受的条件下与eNOS相互作用是否受损，与eNOS功能的缺陷并行。接下来，我们将评估保留eNOS-蛋白质相互作用是否保留eNOS功能和勃起功能。该假设将在用L-4F（载脂蛋白-A-L模拟物）处理的血脂异质小鼠中进行检验，该假设通过维持最佳的eNOS蛋白质相互作用来部分影响eNOS的功能。第三，我们将评估NAD（P）H氧化酶和未偶联的eNOS是否增加了eNOS-蛋白质相互作用和ED在高胆固醇血症小鼠中的ED造成的超氧化物的产生。这将在具有和没有载偶氮蛋白诱导的NAD（P）H氧化酶抑制的高胆固醇小鼠中进行测试。阐明eNOS调控的分子机制对于更好地理解生理和病理生理状况下血管组织中无介导的反应至关重要。拟议的研究有望阐明阴茎中eNOS调节的新分子机制及其在勃起生理学中的生理意义。同时，它可能建议一种特定的eNOS调节途径，作为治疗ED的新分子靶标。