CRT and Mitochondrial Function and Proteome Post-Translational Modifications

CRT 与线粒体功能和蛋白质组翻译后修饰

• 批准号: 8011127
• 负责人: Jennifer E Van Eyk
• 金额: $ 45.57万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8011127
• 关键词: Address; Affect; Agonist; Amino Acids; Antioxidants; Automobile Driving; Basic Science; Biochemical; Cardiac; Cataloging; Catalogs; Clinical; Clinical Treatment; Complex; Computer Simulation; Cyclic AMP-Dependent Protein Kinases; Data; Defect; Enzymes; Equilibrium; Event; Failure; Funding; Heart; Heart Atrium; Heart failure; Individual; Inner mitochondrial membrane; Metabolic; Mitochondria; Mitochondrial Proteins; Mitochondrial Proton-Translocating ATPases; Modeling; Modification; Molecular Weight; Muscle Cells; Myocardial; Oxidation-Reduction; Oxidative Phosphorylation; Oxidative Stress; Pathway interactions; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Post-Translational Protein Processing; Production; Protein Subunits; Proteins; Proteome; Proteomics; Regulation; Research; Respiratory Chain; Respiratory physiology; Role; Site; Structure; Structure-Activity Relationship; Symptoms; Thioredoxin; Transcriptional Regulation; Tyrosine; Ventricular; Work; cell behavior; heart function; improved; improved functioning; molecular phenotype; nitration; oligomycin sensitivity-conferring protein; oxidation; peroxiredoxin; programs; protein complex; protein profiling; research study; respiratory; response; stoichiometry; tool

Cardiac resynchronization therapy (CRT) is an effective clinical heart failure therapy; though how it works at the myocardial level has been largely unknown. An eariy flnding was that CRT improved chamber-level energetic efficiency, but our work during the prior funding period shows a central role for mitochondria and ATP production. DHF and CRT affect the mitochondrial subproteome by speciflcally altering proteins in cellular redox control and oxidative phosphorylation (OxPhos) pathways, manifested by protein quantity and post-translational modiflcations (PTMs) in the mitochondria. For several important targets, such as the mitochondrial ATP synthase (complex V), we have already made fundamental new discoveries about phosphorylation-dependent regulation. Several of the OxPhos complexes can be organized into high molecular weight supercomplexes which can influence global mitochondrial structure and funcflon. This organizaflon is altered in DHF and CRT and seems to be, in part, regulated by speciflc phosphorylaflon events. Additionally, we have found several ROS/RNS-related PTMs occur on mitochondrial proteins and that CRT can blunt these changes, probably by improving the function of antioxidant/scavenger pathways. As such, our underlying hypothesis is that CRT converts the mitochondrial subproteome to a protected phenotype, reversing detrimental DHF-induced protein alterations that regulate key functions to improve ATP production and redox balance. Use of always synchronous failure and an alternative CRT model further strengthens our ability to hone in on changes from resynchronization. Aim 1 focuses on the remodeling of mitochondrial OxPhos protein complexes by CRT which improves the assembly and funcflon of respiratory supercomplexes. Aim 2 invesflgates the modificaflon and regulaflon of mitochondrial proteins by ROS/RNS and their effects on oxidaflon phosphorylation in DHF and CRT hearts. While Aim 3 compares the intramitochondrial phosphorylation of mitochondrial proteins via speciflc responses to activation of PKA, PKC and PKG and their regulaflon during DHF and CRT. In summary. Project 3 uses a large number of proteomic tools to analyze, characterize and quantify the PTMs and pathways behind the CRT-induced improvements to the failing heart, speciflcally focusing on the proteins of the oxidaflve phosphorylaflon subproteome. This data will drive the physiological and biochemical experiments on isolated enzymes and mitochondria which, with the help of computational models of integrated cell behavior, allowing us to further elucidate the underlying molecular phenotype that drives CRT improvements.

心脏再同步治疗（CRT）是一种有效的临床心力衰竭治疗方法；虽然它是如何工作的 心肌水平很大程度上未知。早期发现 CRT 改善了暗室水平 能源效率，但我们在之前的资助期间的工作表明线粒体和 ATP 生产。 DHF 和 CRT 通过特异性改变线粒体中的蛋白质来影响线粒体亚蛋白质组 细胞氧化还原控制和氧化磷酸化 (OxPhos) 途径，通过蛋白质数量和 线粒体中的翻译后修饰 (PTM)。对于几个重要的目标，例如 线粒体 ATP 合酶（复合体 V），我们已经有了关于线粒体 ATP 合酶（复合体 V）的基本新发现 磷酸化依赖性调节。一些 OxPhos 复合物可以组织成高 可以影响整体线粒体结构和功能的分子量超复合物。这 organizaflon 在 DHF 和 CRT 中发生改变，并且似乎部分受到特定的磷酸化 flon 的调节 事件。此外，我们发现一些 ROS/RNS 相关的 PTM 发生在线粒体蛋白上，并且 CRT 可以通过改善抗氧化剂/清除剂途径的功能来减弱这些变化。 因此，我们的基本假设是 CRT 将线粒体亚蛋白质组转化为受保护的蛋白质组。 表型，逆转 DHF 诱导的有害蛋白质改变，调节关键功能以改善 ATP 产生和氧化还原平衡。进一步使用始终同步故障和替代 CRT 模型 增强我们磨练重新同步带来的变化的能力。目标1重点改造 通过 CRT 形成线粒体 OxPhos 蛋白复合物，改善呼吸系统的组装和功能 超级复合物。目标2研究ROS/RNS对线粒体蛋白的修饰和调节 及其对 DHF 和 CRT 心脏中 oxaflon 磷酸化的影响。虽然目标 3 比较了线粒体内 通过对 PKA、PKC 激活的特异性反应来磷酸化线粒体蛋白 PKG 及其在 DHF 和 CRT 期间的调节。总之。项目3使用了大量的 用于分析、表征和量化 CRT 诱导的 PTM 和途径的蛋白质组学工具 改善衰竭的心脏，特别关注氧化磷酰氟的蛋白质 亚蛋白质组。这些数据将推动分离酶的生理和生化实验 线粒体，在整合细胞行为的计算模型的帮助下，使我们能够进一步 阐明推动 CRT 改进的潜在分子表型。