Biochemical Mechanism of Eicosanoid Synthesizing Enzymes

类二十烷酸合成酶的生化机制

• 批准号: 9237609
• 负责人: Aditi Das
• 金额: $ 30.13万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9237609
• 关键词: Acids; Active Sites; Amiodarone; Anti-Inflammatory Agents; Anti-inflammatory; Arachidonic Acids; Binding; Binding Proteins; Biochemical; Biochemistry; CYP2J2 gene; Cardiac Myocytes; Cardiotoxicity; Cardiovascular Diseases; Cardiovascular Physiology; Cardiovascular system; Cholesterol; Complex; Consumption; Detection; Diet; Dietary Fatty Acid; Doxorubicin; Eicosanoids; Electron Transport; Electrostatics; Endocannabinoids; Environment; Enzyme Interaction; Enzymes; Epithelium; Epoxy Compounds; Exhibits; Exogenous Factors; Fluorescence Spectroscopy; Future; Generations; Goals; Health Benefit; Heart; Homeostasis; Human; Inflammation; Kinetics; Link; Linoleic Acids; Lipids; Literature; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Membrane; Membrane Lipids; Metabolism; Methodology; Methods; Muscle Cells; Omega-3 Fatty Acids; Omega-6 Fatty Acids; Outcome; Oxidation-Reduction; Pathway interactions; Pharmaceutical Preparations; Physiological; Play; Positioning Attribute; Process; Proteins; Role; Solubility; Spectrum Analysis; Techniques; Terfenadine; Testing; Thermodynamics; Work; aqueous; cardiovascular health; design; fatty acid metabolism; innovation; lipid mediator; nanodisk; novel; novel strategies; therapeutic development; therapeutic target

PROJECT SUMMARY Dietary consumption of ω-3 and ω-6 fatty acids have been linked to cardiovascular health benefits in humans. The central hypothesis is that the cardiovascular physiological effects of ω-3 and ω-6 fatty acids are partly mediated by the synthesis of eicosanoids via the epoxygenase (EPOX) pathway. Herein we perform biochemical studies of some of the key enzymes in these pathways. CYP2J2 is an enzyme in the EPOX pathway that is highly expressed in the cardiovascular system in the aortic epithelium and cardiomyocytes. CYP2J2's primary effects are facilitated via epoxidation of ω-3 and ω-6 fatty acids into epoxides that exert potent anti- inflammatory, vasodilatory and pro-angiogenic effects. CYP2J2 is also implicated in cardiotoxicity of drugs. Additionally, CYP2J2 is also a membrane bound protein and exhibit unique biochemical mechanisms that are poorly characterized and are the primary focus of the current proposal. Our first goal is to understand allosteric modulation of CYP2J2 epoxygenase activity by ω-3 and ω-6 fatty acids and selected cardiotoxic drugs (doxorubicin, ebastine and terfenadine). Our second goal is to examine the metabolism of ω-3 and ω-6 fatty acid derived endocannabinoids by CYP2J2. It is predicted that similar to ω-6 endocannabinoids, the ω-3 endocannabinoids are substrates for the EPOX enzymes producing novel bioactive epoxide mediators. The third goal is to examine how the composition of membranes effect CYP2J2 activity. We use several novel approaches that includes detection of lipid mediators with mass spectrometry, innovative methodologies such as Nanodiscs to solubilize CYP2J2 and provide membrane bilayer environment. We also introduce novel concepts of lipid-drug heterotropic interactions influencing the formation of the products of these enzymes. The long-term goal of this work is to understand the interplay of the formation of the eicosanoids from dietary fatty acids.

项目摘要 ω-3和ω-6脂肪酸的饮食消费与心血管健康益处有关 人类。中心假设是ω-3和ω-6脂肪酸的心血管生理作用 通过环氧酶（Epox）途径合成类花生酸酯的一部分介导。在这里我们 对这些途径中一些关键酶进行生化研究。 CYP2J2是一种酶 在主动脉上皮的心血管系统中高度表达的环氧途径和 心肌细胞。 CYP2J2的主要作用是通过将ω-3和ω-6脂肪酸的环氧化制备的 发挥潜在抗炎，血管舒张和促血管生成作用的环氧化物。 CYP2J2也是 在药物的心脏毒性中实施。此外，CYP2J2也是一种膜结合蛋白，并且展示 特征不佳的独特生化机制，是电流的主要重点 提议。我们的第一个目标是了解ω-3对CYP2J2环氧酶活性的变构调制 和ω-6脂肪酸以及选定的心脏毒性药物（阿霉素，ebastine和terfenadine）。我们的第二个 目标是检查CYP2J2衍生的内源性大麻素的代谢。这是 预测与ω-6内源性大麻素类似，ω-3内源性大麻素是环氧氧基的底物 产生新型生物活性环氧介质的酶。第三个目标是检查构图如何 机制影响CYP2J2活性。我们使用几种新方法，包括检测脂质 具有质谱，创新方法（例如纳米盘）的介体 并提供膜双层环境。我们还介绍了脂质毒物异型的新颖概念 相互作用会影响这些酶产物的形成。这项工作的长期目标 是为了了解饮食脂肪酸形成类eic烷的相互作用。