CRT and Mitochondrial Function and Proteome Post-Translational Modifications

心脏复律治疗（CRT）是一种有效的临床心力衰竭治疗方法，但它是如何工作的， 心肌水平在很大程度上是未知的。早期发现CRT改善了室平面 能量效率，但我们在前一个资助期间的工作显示了线粒体的核心作用， ATP生产。DHF和CRT通过特异性改变线粒体蛋白质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，表现为蛋白质数量和 翻译后修饰（PTM）在线粒体。对于几个重要目标，例如 线粒体ATP合酶（复合物V），我们已经有了关于 磷酸化依赖性调节。几种OxPhos络合物可以被组织成高分子量的。 分子量超复合物，可以影响整体线粒体结构和功能。这 在DHF和CRT中，organizaflon发生改变，似乎部分受特异性磷酸化酶调节 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， CRT可能通过改善抗氧化剂/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，逆转有害的DHF诱导的蛋白质改变，调节关键功能，以改善 ATP生成和氧化还原平衡。始终同步故障和替代CRT模型的使用还 增强了我们从信息化中磨练变化的能力。目标1的重点是重塑 线粒体OxPhos蛋白复合物通过CRT改善呼吸道的组装和功能 超复合体目的2研究ROS/RNS对线粒体蛋白质的修饰和调节 以及它们对DHF和CRT心脏中的oxidaflon磷酸化的影响。虽然目标3比较了线粒体内 通过对PKA、PKC激活的特异性反应使线粒体蛋白磷酸化 和PKG及其在DHF和CRT期间的调节。总之。项目3使用大量的 蛋白质组学工具，以分析，表征和量化PTM和CRT诱导的 改善衰竭的心脏，特别是关注氧化磷脂酰肌醇的蛋白质 亚蛋白质组这些数据将推动对分离酶的生理和生化实验， 线粒体，在综合细胞行为的计算模型的帮助下，使我们能够进一步 阐明驱动CRT改善的潜在分子表型。

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.