CRT and Mitochondrial Function and Proteome Post-Translational Modifications

心脏再同步化治疗(CRT)是一种有效的临床心力衰竭治疗方法； 心肌水平在很大程度上是未知的。一个早期的迹象是CRT提高了燃烧室的水平 能量效率，但我们在前一个资助期的工作表明线粒体和 ATP生产。DHF和CRT通过特异地改变线粒体亚蛋白质组中的蛋白质 细胞氧化还原控制和氧化磷酸化(OxPhos)途径，表现为蛋白质和 线粒体的翻译后修饰(PTM)。对于几个重要的目标，例如 线粒体ATP合成酶(复合体V)，我们已经有了关于 依赖于磷酸化的调节。几个OxPhos络合物可以被组织成高 可影响全球线粒体结构和功能的分子量超复合体。这 在DHF和CRT中，Organaflon被改变，并且似乎在一定程度上受特定的磷酸化aflon调节 事件。此外，我们还发现一些与ROS/RNS相关的PTM存在于线粒体蛋白和 CRT可以钝化这些变化，可能是通过改善抗氧化剂/清道夫途径的功能。 因此，我们的基本假设是，CRT将线粒体亚蛋白质组转换为受保护的 表型，逆转DHF诱导的调节关键功能以改善的有害蛋白质变化 ATP生产和氧化还原平衡。使用始终同步故障和替代CRT模型进一步 增强了我们从重新同步中磨练变化的能力。目标1专注于重塑 CRT制备线粒体OxPhos蛋白复合体改善呼吸系统的组装和功能 超复合体。目的2研究ROS/RNS对线粒体蛋白质的修饰和调节作用 以及它们对DHF和CRT心脏中氧化法酮磷酸化的影响。而Aim 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中的调节作用。总而言之。项目3使用了大量的 蛋白质组学工具分析、表征和量化CRT诱导的PTM和途径 对衰竭心脏的改善，特别是关注氧化磷脂的蛋白质 亚蛋白质组。这一数据将推动对分离的酶和 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改进的潜在分子表型。

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.