Pin1 prolyl isomerase regulates endothelial nitric oxide synthase

Pin1 脯氨酰异构酶调节内皮一氧化氮合酶

基本信息

批准号：
8823817
负责人：
RICHARD C VENEMA
金额：
$ 36.94万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2016-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8823817
关键词：
Acute Adhesions Affect Agonist Alzheimer&apos s Disease Amino Acid Sequence Animal Disease Models Animal Model Animals Apoptosis Arginine Arterial Fatty Streak Atherosclerosis Binding Blood Platelets Blood Pressure Blood Vessels Cardiovascular Diseases Cardiovascular system Cattle Clinical Trials Complex Data Development Diabetes Mellitus Disease Endothelial Cells Endothelium Enzymes Exposure to Extravasation Functional disorder Goals Graft Rejection Homeostasis Hypertension In Vitro Inflammation Inflammatory Investigation Knowledge Lead Leukocytes Maintenance Malignant Neoplasms Methods Mitogen Activated Protein Kinase 1 Mitogen-Activated Protein Kinase 3 Nitric Oxide Organ Transplantation Pathogenesis Peptidylprolyl Isomerase Phosphorylation Phosphorylation Site Physiology Platelet aggregation Post-Translational Regulation Process Production Proline Proline-Directed Protein Kinases Protein Conformation Proteins Publishing Regulation Research Proposals Role Scientist Serine Site Structure Testing Threonine Time Vascular Diseases Vasodilation angiogenesis atherogenesis blood pressure regulation cardiovascular health cytokine disease phenotype human NOS3 protein in vivo inhibitor/antagonist interest member novel oxidation prevent protein function protein protein interaction vascular smooth muscle cell proliferation

项目摘要

DESCRIPTION (provided by applicant): Endothelium-derived nitric oxide (NO) regulates many important aspects of cardiovascular homeostasis and alterations in vascular NO production of as little as two-fold can have major consequences in both normal vascular physiology and vascular pathophysiology. NO has a crucial role in regulation of the state of vasodilatation of blood vessels and hence in regulation of blood pressure. NO released from the endothelium also modulates other processes as well including platelet aggregation, platelet and leukocyte adhesion to the endothelium, vascular smooth muscle cell proliferation, endothelial cell apoptosis, angiogenesis, and vascular leakage involved in acute inflammation. Because of the key role of NO in each of these processes, abnormalities in vascular NO production are thought to contribute to the pathogenesis of certain vascular disorders such as those of atherosclerosis, diabetes, and hypertension. NO is synthesized in endothelial cells by oxidation of arginine catalyzed by the enzyme, endothelial nitric oxide synthase (eNOS). Obtaining a detailed understanding of the structure, function, and regulation of eNOS enzyme is thus clearly a worthy goal of scientific investigation. eNOS is regulated post-translationally by two primary mechanisms: protein-protein interactions and phosphorylation. Nothing is yet known about how these two different eNOS regulation processes are interrelated. What has been known for some time, however, is that there exists in eNOS an inhibitory phosphorylation site at serine 116 (S116). This site is phosphorylated in endothelial cells under basal conditions and after exposure to pro-inflammatory cytokines. Our preliminary data give rise to the hypothesis that proline-directed phosphorylation of eNOS at S116 promotes binding of the Pin1 prolyl isomerase and Pin1-induced conformational changes that suppress eNOS activity. Preliminary data also show that this process may make an important contribution to the disease of atherosclerosis. We therefore propose to test these hypotheses by a variety of different methods using purified proteins, cultured endothelial cells, intact blood vessel segments, and whole animals including animal disease models. These studies will provide a more complete understanding than currently exists of regulation of eNOS in endothelial cells, a topic of great interest to both basic scientists and clinicians.

描述（由申请人提供）：内皮源性一氧化氮 (NO) 调节心血管稳态的许多重要方面，血管一氧化氮生成的改变只要两倍即可对正常血管生理学和血管病理生理学产生重大影响。一氧化氮在血管舒张状态的调节以及血压的调节中起着至关重要的作用。内皮释放的一氧化氮还调节其他过程，包括血小板聚集、血小板和白细胞与内皮的粘附、血管平滑肌细胞增殖、内皮细胞凋亡、血管生成和急性炎症中涉及的血管渗漏。由于一氧化氮在这些过程中的每一个过程中都发挥着关键作用，因此血管一氧化氮生成异常被认为是导致某些血管疾病（例如动脉粥样硬化、糖尿病和高血压）的发病机制的原因之一。 NO 在内皮细胞中通过内皮一氧化氮合酶 (eNOS) 催化精氨酸氧化而合成。因此，详细了解 eNOS 酶的结构、功能和调节显然是一个值得科学研究的目标。 eNOS 通过两种主要机制进行翻译后调节：蛋白质-蛋白质相互作用和磷酸化。目前尚不清楚这两种不同的 eNOS 调节过程如何相互关联。然而，一段时间以来人们已经知道，eNOS 中的丝氨酸 116 (S116) 存在一个抑制性磷酸化位点。在基础条件下和暴露于促炎细胞因子后，该位点在内皮细胞中被磷酸化。我们的初步数据提出了这样的假设：eNOS 在 S116 处的脯氨酸定向磷酸化促进 Pin1 脯氨酰异构酶的结合以及 Pin1 诱导的构象变化，从而抑制 eNOS 活性。初步数据还表明，这一过程可能对动脉粥样硬化疾病做出重要贡献。因此，我们建议使用纯化的蛋白质、培养的内皮细胞、完整的血管片段和包括动物疾病模型在内的整个动物，通过各种不同的方法来检验这些假设。这些研究将比目前对内皮细胞中 eNOS 的调节提供更全面的了解，这是基础科学家和临床医生都非常感兴趣的话题。