Regulation of Mitochondrial Metabolism by Post-Translational Modifications

翻译后修饰对线粒体代谢的调节

基本信息

批准号：
9262822
负责人：
David J Pagliarini
金额：
$ 30.16万
依托单位：
MORGRIDGE INSTITUTE FOR RESEARCH, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-15 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9262822
关键词：
3-hydroxy-3-methylglutaryl-coenzyme A Acetylation Acyl CoA Dehydrogenases Address Affect Age Aging Algorithms Biochemistry Bioenergetics Biological Assay Cells Data Development Disease Employee Strikes Enzymes Eukaryotic Cell Event Fatty Acids Foundations Functional disorder Future Goals Heart Diseases In Vitro Inborn Errors of Metabolism Knowledge Liver Liver Mitochondria Malignant Neoplasms Maps Mass Spectrum Analysis Measures Medicine Metabolic Diseases Metabolic Pathway Metabolism Mitochondria Mitochondrial Proteins Modification Mouse Strains Non-Insulin-Dependent Diabetes Mellitus Obesity Organelles Pathogenicity Peptides Phosphorylation Phosphorylation Site Phosphotransferases Physiology Post-Translational Protein Processing Process Protein Acetylation Protein Kinase Proteins Proteomics Regulation Reproducibility Role Sampling Series Signal Transduction Signaling Protein Site Techniques Testing Therapeutic Therapeutic Intervention activity-based protein profiling acyl-CoA dehydrogenase base cell type data acquisition enzyme activity human disease infancy innovation insight ketogenesis metabolomics mitochondrial dysfunction mitochondrial metabolism mouse model new therapeutic target novel novel therapeutic intervention novel therapeutics oxidation phosphoproteomics prevent protein function public health relevance tool

项目摘要

DESCRIPTION (provided by applicant): Mitochondria are centers of metabolism and signaling whose function is essential to all but a few eukaryotic cell types. General dysfunction of these organelles is implicated in a wide range of inborn errors of metabolism, and in an increasing number of common human diseases, including type 2 diabetes (T2D), cancer and heart disease. However, the specific alterations that underlie mitochondrial dysfunction in these disorders are most often poorly defined, and are nearly always impervious to therapeutic intervention. As such, clearly defining the pathogenic mitochondrial alterations that underlie metabolic disorders and devising new therapeutic strategies to treat these conditions represent principal challenges in mitochondrial medicine. Emerging evidence, including our own recent proteomic data, have revealed that mitochondrial proteins are replete with phosphorylation and acetylation sites that change dynamically between healthy and diseased states, and that these modifications are frequently found jointly on the same proteins. These data suggest that these post- translational modifications (PTMs) are widely important in regulating mitochondrial metabolism, and that aberrant levels of these modifications are among the relevant alterations underlying mitochondrial pathophysiology. If true, this could motivate the development of a novel therapeutic strategy: the control of mitochondrial metabolism via manipulation of cellular signaling processes. However, despite these promising early studies on mitochondrial PTMs, our understanding of their role in mitochondrial physiology remains in its infancy for a number of reasons. First, it remains unclear which of these modifications are actually important for regulating protein function. Second, there is a striking lack of information regarding the enzymes (e.g., kinases) that perform these modifications. These substantial knowledge gaps prevent us from understanding how cells use PTMs to manipulate mitochondrial function, and from exploiting this information for potential therapeutic benefit. This proposal takes a thorough and innovative approach to addressing these knowledge gaps by blending focused biochemistry with a range of state-of-the-art mass spectrometry tools. In particular, the overarching goals of this proposal are to 1) elucidate how phosphorylation affects the activities of select liver mitochondrial proteins involved in ketogenesis and beta-oxidation using in vitro biochemistry and cell-based metabolomics and bioenergetics, 2) to identify kinases in mitochondria that execute these events through activity-based and targeted mass spectrometry techniques, and 3) to establish a complementary, quantitative map of the dynamic mitochondrial protein acetylation events that accompany the onset of obesity and T2D. Completion of these aims will provide new insight into the regulation of key mitochondrial metabolic pathways, will provide a foundation for future mechanistic studies into the interrelationship between mitochondrial post-translational modifications, and will help reveal new therapeutic targets for the treatment of mitochondrial dysfunction.

描述（由申请人提供）：线粒体是代谢和信号传导的中心，其功能对于除少数真核细胞类型之外的所有真核细胞类型都是必需的。这些细胞器的一般功能障碍与多种先天性代谢错误以及越来越多的常见人类疾病有关，包括 2 型糖尿病 (T2D)、癌症和心脏病。然而，这些疾病中线粒体功能障碍的具体改变通常是不清楚的，并且几乎总是不受治疗干预的影响。因此，明确界定代谢紊乱背后的致病性线粒体改变并设计新的治疗策略来治疗这些疾病是线粒体医学的主要挑战。新出现的证据，包括我们最近的蛋白质组学数据，表明线粒体蛋白质充满了在健康和患病状态之间动态变化的磷酸化和乙酰化位点，并且这些修饰经常同时出现在相同的蛋白质上。这些数据表明，这些翻译后修饰（PTM）对于调节线粒体代谢非常重要，并且这些修饰的异常水平是线粒体病理生理学基础的相关改变之一。如果这是真的，这可能会激发一种新的治疗策略的开发：通过操纵细胞信号传导过程来控制线粒体代谢。然而，尽管这些关于线粒体 PTM 的早期研究很有希望，但由于多种原因，我们对其在线粒体生理学中的作用的理解仍处于起步阶段。首先，目前尚不清楚哪些修饰对于调节蛋白质功能实际上很重要。其次，关于执行这些修饰的酶（例如激酶）的信息明显缺乏。这些巨大的知识差距使我们无法理解细胞如何利用 PTM 来操纵线粒体功能，也无法利用这些信息来获得潜在的治疗益处。该提案采用彻底和创新的方法，通过将重点生物化学与一系列最先进的质谱工具相结合来解决这些知识差距。特别是，该提案的总体目标是 1) 使用体外生物化学和基于细胞的代谢组学和生物能量学阐明磷酸化如何影响参与生酮和 β-氧化的精选肝线粒体蛋白的活性，2) 识别线粒体中的激酶通过基于活动和有针对性的质谱技术执行这些事件，3) 建立伴随肥胖和肥胖发作的动态线粒体蛋白乙酰化事件的补充定量图谱T2D。这些目标的完成将为关键线粒体代谢途径的调节提供新的见解，将为未来线粒体翻译后修饰之间相互关系的机制研究奠定基础，并将有助于揭示治疗线粒体功能障碍的新治疗靶点。