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

基本信息

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
项目类别：
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

项目摘要

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.

心脏重新同步治疗（CRT）是一种有效的临床心力衰竭疗法；虽然它如何工作心肌水平在很大程度上是未知的。 CRT改善了室级别充满活力的效率，但是我们在上一期间的工作表明了线粒体和 ATP生产。 DHF和CRT通过特异性改变蛋白质的蛋白质影响线粒体亚蛋白细胞氧化还原对照和氧化磷酸化（OXPHOS）途径，由蛋白质数量和线粒体中的翻译后修饰（PTM）。对于几个重要目标，例如线粒体ATP合酶（复杂V），我们已经做出了有关的基本新发现磷酸化依赖性调节。可以将几种Oxphos配合物组织成高分子量超复合物可以影响整体线粒体结构和构氟。这组织中有DHF和CRT改变了组织，似乎部分地受到peciflc Phosphorylaflon的调节事件。此外，我们发现线粒体蛋白上发生了几个与ROS/RN相关的PTM，并且 CRT可以通过改善抗氧化剂/清除途径的功能来钝化这些变化。因此，我们的基本假设是CRT将线粒体亚蛋白质转换为受保护表型，逆转有害DHF诱导的蛋白质改变，这些蛋白质改变了关键功能以改善 ATP生产和氧化还原平衡。始终使用同步故障和替代CRT模型增强了我们磨练重新同步变化的能力。 AIM 1专注于重塑 CRT的线粒体Oxphos蛋白复合物改善了呼吸系统的组装和功能超复合。 AIM 2 Invesflates通过ROS/RNS的线粒体蛋白的修饰和质量以及它们对DHF和CRT心脏中氧化磷酸化的影响。而AIM 3比较了Intibochondirial 线粒体蛋白通过PKA激活PKC的激活反应对线粒体蛋白的磷酸化以及DHF和CRT期间的PKG及其Regulaflon。总之。项目3使用大量分析，表征和量化CRT诱导的PTM和途径的蛋白质组学工具改善失败的心脏，专门针对氧化氟氟氟烷的蛋白质次蛋白酶。这些数据将驱动孤立酶的生理和生化实验线粒体，借助集成细胞行为的计算模型，使我们能够进一步阐明驱动CRT改进的基本分子表型。