The Role of Protease-Activated Receptors in the Regulation of eNOS

蛋白酶激活受体在 eNOS 调节中的作用

基本信息

批准号：
8021791
负责人：
Evangeline D Motley-Johnson
金额：
$ 10.88万
依托单位：
MEHARRY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-01 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8021791
关键词：
ADRBK1 gene Address Adhesions African American Agonist Americas Area Bacteriophages Biological Assay Biology Blood Vessels Calmodulin Cardiovascular Diseases Cardiovascular system Cattle Cause of Death Cell Proliferation Cell physiology Cells Chimera organism Chimeric Proteins Communities Couples Coupling Development Disease Doctor of Philosophy Dominant-Negative Mutation Endothelial Cells Endothelium Enzymes Etiology F2R gene Family Functional disorder Funding G-Protein-Coupled Receptors GTP-Binding Proteins Goals Grant Health Heterotrimeric GTP-Binding Proteins Hypertension In Vitro Laboratories Lead Leukocytes Measures Mediating Minority Molecular Mutation Nitric Oxide Paper Peptide Hydrolases Permeability Phage Display Pharmaceutical Preparations Phosphorylation Phosphotransferases Physiology Pilot Projects Play Production Protein Isoforms Protein Kinase Inhibitors Proteinase-Activated Receptors Proteins Proteomics Public Health Publications Publishing RGS Domain Receptor Activation Receptor Signaling Regulation Relaxation Research Rho-associated kinase Risk Factors Role Schools Science Scientist Signal Pathway Signal Transduction Signal Transduction Pathway Site Structure Structure-Activity Relationship Students Tail Techniques Testing Therapeutic Thrombin Training Translating Underserved Population angiogenesis career cell motility health disparity human NOS3 protein inhibitor/antagonist interest mutant novel prevent professor protein activation protein kinase C-delta protein kinase inhibitor receptor research study response rho tool treatment strategy vector

项目摘要

DESCRIPTION (provided by applicant): The objective of this study is to explore the signal transduction mechanism by which protease-activated receptors (PARs) regulate endothelial nitric oxide synthase (eNOS) phoshorylation and activation in endothelial cells. Proteases play very important roles in endothelial physiology and pathophysiology through activation of PARs. Their functions include regulations of endothelium-dependent contraction/relaxation, endothelial permeability/barrier function, leukocyte adhesion, angiogenesis, endothelial cell migration and proliferation. Thus, PARs are very likely to be involved in the development of cardiovascular diseases. However, the detailed molecular mechanisms by which PARs regulate these functions remain largely unclear. PAR1 and PAR2 are believed to play major roles in endothelial cell function. Recent studies revealed multiple heterotrimeric G protein-coupling (Gq, Gi, G12/13) and several downstream signaling cascades of PAR1 in endothelial cells and other cells. By contrast, much less is known regarding PAR2 signal transduction in endothelial cells, which mainly couples to Gq. Our goal is to study the specific signal transduction pathways utilized by PARs including heterotrimeric G protein- coupling together with the structure-activity relationship of PARs during coupling and their roles in mediating downstream signals that regulate eNOS. We hypothesize that PAR1 and PAR2 reciprocally regulate eNOS through distinct signaling pathways and have proposed the following specific aims to address this hypothesis. We plan: 1. To demonstrate that a distinct receptor C-tail structure associated with distinct G protein-coupling is required for reciprocal eNOS phosphorylation by PARs. 2. To identify eNOS kinases that reciprocally regulate eNOS activity in response to distinct PARs. 3. To identify the distinct PAR C-tail associated proteins that are responsible for reciprocal eNOS regulation. Cultured endothelial cells will be used to detect phosphorylation of eNOS. Kinase assays, site mutations, and novel proteomic approaches will be used to identify other proteins involved in the reciprocal phosphorylation of eNOS. The results of this study will be valuable in developing specific-acting therapeutic drugs that can be used to treat endothelial dysfunctions that lead to cardiovascular diseases. PUBLIC HEALTH RELEVANCE: This project is relevant to public health because of its importance in understanding some of the possible causes of cardiovascular disease, the number one killer in America, especially among the African American community. It will provide information regarding the role of protease-activated receptors in the regulation of the endothelium, which plays an important role in vascular tone. Endothelial dysfunction is a major cause of hypertension, and these studies will contribute to the development of treatment strategies to alleviate or prevent cardiovascular disease.

描述（申请人提供）：本研究的目的是探讨蛋白酶激活受体（PAR）调节内皮细胞中内皮一氧化氮合酶（eNOS）磷酸化和激活的信号转导机制。蛋白酶通过激活 PAR，在内皮生理学和病理生理学中发挥非常重要的作用。它们的功能包括调节内皮依赖性收缩/舒张、内皮通透性/屏障功能、白细胞粘附、血管生成、内皮细胞迁移和增殖。因此，PAR很可能参与心血管疾病的发生发展。然而，PAR 调节这些功能的详细分子机制仍不清楚。 PAR1 和 PAR2 被认为在内皮细胞功能中发挥重要作用。最近的研究揭示了内皮细胞和其他细胞中 PAR1 的多个异源三聚体 G 蛋白偶联（Gq、Gi、G12/13）和几个下游信号级联。相比之下，人们对内皮细胞中的 PAR2 信号转导知之甚少，该信号主要与 Gq 偶联。我们的目标是研究 PAR 使用的特定信号转导途径，包括异三聚体 G 蛋白偶联以及偶联过程中 PAR 的构效关系以及它们在介导调节 eNOS 的下游信号中的作用。我们假设 PAR1 和 PAR2 通过不同的信号通路相互调节 eNOS，并提出以下具体目标来解决这一假设。我们计划： 1. 证明 PAR 相互磷酸化 eNOS 需要与不同 G 蛋白偶联相关的独特受体 C 尾结构。 2. 鉴定响应不同 PAR 相互调节 eNOS 活性的 eNOS 激酶。 3. 鉴定负责 eNOS 相互调节的不同 PAR C 尾相关蛋白。培养的内皮细胞将用于检测 eNOS 的磷酸化。激酶测定、位点突变和新的蛋白质组学方法将用于鉴定参与 eNOS 相互磷酸化的其他蛋白质。这项研究的结果对于开发可用于治疗导致心血管疾病的内皮功能障碍的特异性治疗药物具有重要意义。公共卫生相关性：该项目与公共卫生相关，因为它对于了解心血管疾病的一些可能原因很重要，心血管疾病是美国的头号杀手，特别是在非裔美国人社区中。它将提供有关蛋白酶激活受体在内皮调节中的作用的信息，内皮在血管张力中发挥重要作用。内皮功能障碍是高血压的主要原因，这些研究将有助于制定缓解或预防心血管疾病的治疗策略。