Systems-to-structure approaches for defining mitochondrial protein function

定义线粒体蛋白质功能的系统到结构方法

• 批准号: 10370341
• 负责人: David J Pagliarini
• 金额: $ 64.58万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370341
• 关键词: Address; Alzheimer' s Disease; Anabolism; Area; Automobile Driving; Binding Proteins; Biochemical; Biochemistry; Bioenergetics; Biogenesis; Biology; Cells; Cellular biology; Custom; Disease; Eukaryotic Cell; Functional disorder; Genetic Transcription; Goals; Human; Inborn Errors of Metabolism; Infrastructure; Knowledge; Lipid Binding; Mass Spectrum Analysis; Metabolism; Mitochondria; Mitochondrial Proteins; Nature; Non-Insulin-Dependent Diabetes Mellitus; Organelles; Orphan; Parkinson Disease; Pathway interactions; Phosphotransferases; Plant Roots; Positioning Attribute; Process; Proteins; Proteome; Research; Role; Signal Transduction; Structure; System; Therapeutic; Ubiquinone; Yeasts; base; cancer type; cell type; design; human disease; knockout gene; meetings; mitochondrial dysfunction; novel; novel therapeutic intervention; programs; protein function; protein protein interaction; systematic biology

PROJECT SUMMARY Mitochondria are centers of metabolism and signaling whose function is essential to all but a few eukaryotic cell types. Despite their position as the iconic powerhouses of cellular biology, many aspects of mitochondria remain remarkably obscure—a fact that contributes to our near complete inability to address mitochondrial dysfunction therapeutically. Such dysfunction is associated with a spectrum of rare inborn errors of metabolism and an increasing number of common diseases—including Parkinson’s, Alzheimer’s, various cancers, and type 2 diabetes—often through distinct means. For instance, aberrant mitochondrial biogenesis can fail to properly set cellular mitochondrial content; dysregulated signaling processes can fail to calibrate mitochondrial activity to changing cellular needs; and malfunctioning proteins can render core bioenergetic processes ineffectual. A major bottleneck to understanding—and ultimately addressing—these processes is that the proteins driving them have often not been identified. Concurrently, the functions of hundreds of known mitochondrial proteins that may fulfill these roles are undefined, or at best are poorly understood. In 2008, I led an integrative effort to generate a comprehensive compendium of the mammalian mitochondrial proteome—termed MitoCarta—that doubled the number of known mammalian mitochondrial proteins and exposed this major gap in knowledge: A striking ~300 of the ~1100 proteins had no annotated function, including ~50 that are now directly associated with human disease. Thus, the high-level goal of my research program is to achieve a more comprehensive understanding of mitochondrial biology by systematically establishing the functions of orphan mitochondrial proteins and their roles within disease-related processes. We do so by first devising novel, multi-dimensional analyses designed to make new connections between these proteins and established pathways and processes. These include customized, high-throughput protein-protein interaction screens, large- scale mass spectrometry-based profiling of yeast and human cell gene knockouts, and computational approaches. We then employ mechanistic and structural approaches to define the functions of select proteins at biochemical depth, including ancient and atypical kinases and lipid binding proteins that enable the mitochondrial coenzyme Q biosynthesis pathway and other essential metabolic processes. Finally, we investigate how post- transcriptional and post-translational regulators operate to establish a customized mitochondrial infrastructure capable of meeting changing cellular needs. Overall, by purposefully elucidating the unexplored areas of mitochondrial biology, we are rapidly arriving at a more complete understanding of what these organelles do and how their protein componentry enables their myriad functions. These efforts promise to help establish a deep, mechanistic understanding of mitochondrial biochemistry that will motivate novel therapeutic strategies for the vast array of human disorders rooted in mitochondrial dysfunction. !

项目总结 线粒体是新陈代谢和信号传递的中心，其功能对除少数真核细胞外的所有真核细胞都是必不可少的。 类型。尽管它们是细胞生物学的标志性动力源，但线粒体的许多方面仍然存在 令人费解--这一事实导致我们几乎完全无法解决线粒体功能障碍 从治疗上讲。这种功能障碍与一系列罕见的先天性新陈代谢错误和 越来越多的常见疾病--包括帕金森氏症、阿尔茨海默氏症、各种癌症和2型 糖尿病--通常是通过不同的方式。例如，异常的线粒体生物发生可能无法正确设置 细胞线粒体含量；失调的信号传递过程可能无法校准线粒体的活性 改变细胞需求；以及功能失调的蛋白质可能使核心生物能量过程无效。一位少校 理解并最终解决这些过程的瓶颈是驱动它们的蛋白质 通常不会被发现。同时，数百种已知的线粒体蛋白的功能可能实现 这些角色没有定义，或者充其量也就是人们对它们知之甚少。2008年，我领导了一项综合努力，以产生 哺乳动物线粒体蛋白质组的综合概要-称为MitoCarta-将 已知的哺乳动物线粒体蛋白的数量，并暴露了这一重大知识差距：惊人的~300 其中~1100个蛋白质没有注释功能，包括现在与人类直接相关的~50个蛋白质 疾病。因此，我的研究计划的高层次目标是实现更全面的 通过系统确定孤儿的功能来理解线粒体生物学 线粒体蛋白及其在疾病相关过程中的作用。我们通过首先设计小说来做到这一点， 多维分析旨在建立这些蛋白质之间的新联系 途径和过程。这些包括定制的高通量蛋白质-蛋白质相互作用屏幕，大型 基于尺度质谱学的酵母和人类细胞基因敲除的图谱，以及计算 接近了。然后，我们使用机械论和结构方法来定义选定蛋白质的功能 生化深度，包括古老的和非典型的激酶和脂结合蛋白，使线粒体 辅酶Q的生物合成途径和其他必要的代谢过程。最后，我们调查了POST如何 转录和翻译后调节因子的作用是建立定制的线粒体基础结构 能够满足不断变化的蜂窝需求。总体而言，通过有目的地阐明 线粒体生物学，我们正在迅速对这些细胞器的功能和功能有更全面的了解 它们的蛋白质成分是如何使它们具有多种功能的。这些努力承诺帮助建立一个深层次的、 对线粒体生物化学的机械性理解将激励新的治疗策略 大量的人类疾病根源于线粒体功能障碍。 好了！