Aging Mitochondrial Interactome

衰老线粒体相互作用组

基本信息

批准号：
10688325
负责人：
James Edward Bruce
金额：
$ 36.14万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-30 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10688325
关键词：
ATP Synthesis Pathway Activity Cycles Address Age Aging Biological Assay Biopsy Brain Cardiovascular system Chemicals Citric Acid Cycle Complex Data Elderly Electron Transport Functional disorder Heart Mitochondria Homeostasis Human Hydrogen Peroxide In Vitro Individual Intervention Kidney Lead Maps Mass Spectrum Analysis Measures Mediating Membrane Potentials Mitochondria Mitochondrial Proteins Molecular Mus Muscle Muscle Mitochondria Muscle function Myocardium Nature Organelles Oxidants Oxidation-Reduction Oxidative Stress Paraquat Pathology Pathway interactions Peptides Permeability Phosphorylation Play Positioning Attribute Production Protein Conformation Proteins Quality of life Reactive Oxygen Species Research Respiration Role Site Skeletal Muscle Stress System Testing Tissues Translating Translations Uncertainty Vision age related aged body system catalase complex IV crosslink experimental study functional decline heart function human tissue improved in vivo innovation insight mitochondrial dysfunction mitochondrial metabolism mouse model oligomycin sensitivity-conferring protein protein complex public health relevance resilience tool young adult

项目摘要

Changes in mitochondrial function play a central role in age-related pathologies, loss of resilience, and the decline in quality of life in older adults. As we age there is a shift in our mitochondria toward a reduced ability to generate ATP and increased oxidant production. These changes lead to disruption of redox and energy homeostasis, altered metabolite levels, and increased sensitivity to permeability transition, all of which contribute to tissue dysfunction. Mitochondria are dynamic organelles that continuously adapt to changing cellular demands by altering protein assembly and interactions to modify their function. Despite the obvious importance, little is known about how age-related changes in mitochondrial protein interactions (interactome) underlie changes in function with age. To address this fundamental question, we propose to apply a state of the art quantitative chemical cross-linking with mass spectrometry (qXL-MS) strategy to quantify changes in the mitochondrial interactome with age. By combining this innovative qXL-MS approach with detailed assays of mitochondrial metabolism and interventions we have developed over the last several years to manipulate mitochondria in vivo and in vitro, we are uniquely positioned to identify the molecular level changes in mitochondrial interactome that underlie age-related mitochondrial dysfunction. Our preliminary data indicate disruption of multiple protein interaction networks involved in ADP transport, ATP synthesis, and substrate supply to the electron transport system in aged heart and skeletal muscle. These changes are associated with previously demonstrated decreases in ATP production and lower sensitivity to ADP. Furthermore, we have shown that a mitochondrial targeted intervention (SS-31) that reverses mitochondrial dysfunction in heart and skeletal muscle, specifically interacts with many of the same protein complexes that our interactome studies reveal are disrupted in aging, including the ANT and complexes IV and V of the electron transport system. Our overall hypothesis tested in this proposal is that changes in the mitochondrial interactome with age underlie decreased ATP production and increased oxidant production in mitochondria from aged heart and skeletal muscle. Aim 1 applies XL-MS and protein and site-specific mitochondrial assays to quantify changes in the mitochondrial interactome with age and those induced by pro-oxidant treatment in vivo and in vitro in mouse heart and skeletal muscle. Aim 2 quantifies the effect on the mitochondrial interactome of two well established mitochondrial targeted interventions, SS-31 and mitochondrial targeted catalase, to identify the most important age-related changes in the protein interaction networks. Aim 3 tests whether changes in the mitochondrial interactome identified in aims 1 and 2 translate into aged human skeletal muscle. The mitochondrial interactome, function, and effects of SS-31 from older adults separated into low and high performing groups are compared with young adults. Results from these experiments will have a significant impact and have the potential to transform the field by providing the first test of how disruption of mitochondrial protein interactions contribute to age-related mitochondrial dysfunction. 1

线粒体功能的变化在与年龄相关的病理、恢复力丧失和老年人的生活质量下降。随着年龄的增长，我们的线粒体的能力会降低产生 ATP 并增加氧化剂的产生。这些变化导致氧化还原和能量的破坏体内平衡、代谢物水平改变以及对渗透性转变的敏感性增加，所有这些都有助于导致组织功能障碍。线粒体是动态细胞器，不断适应不断变化的细胞需求通过改变蛋白质组装和相互作用来修改其功能。尽管重要性显而易见，但很少有了解与年龄相关的线粒体蛋白质相互作用（相互作用组）的变化如何导致功能随年龄增长。为了解决这个基本问题，我们建议应用最先进的定量方法化学交联与质谱（qXL-MS）策略来量化线粒体的变化与年龄的相互作用。通过将这种创新的 qXL-MS 方法与线粒体的详细分析相结合我们在过去几年中开发的代谢和干预措施，用于在体内操纵线粒体在体外，我们具有独特的优势来识别线粒体相互作用组的分子水平变化，是与年龄相关的线粒体功能障碍的基础。我们的初步数据表明多种蛋白质被破坏参与 ADP 传输、ATP 合成和电子传输底物供应的相互作用网络衰老的心脏和骨骼肌系统。这些变化与之前证明的相关 ATP 产生减少，对 ADP 的敏感性降低。此外，我们还发现线粒体定向干预（SS-31）可逆转心脏和骨骼肌的线粒体功能障碍，特别是与许多相同的蛋白质复合物相互作用，我们的相互作用组研究表明，这些蛋白质复合物在衰老过程中受到破坏，包括ANT和电子传递系统的复合物IV和V。我们的总体假设在此得到检验推测线粒体相互作用组随年龄的变化是 ATP 产生减少的基础衰老心脏和骨骼肌线粒体中氧化剂的产生增加。目标 1 应用 XL-MS 和蛋白质和位点特异性线粒体测定，以量化线粒体相互作用组随年龄和年龄的变化在小鼠心脏和骨骼肌中通过体内和体外促氧化处理诱导的那些。目标 2 量化两种成熟的线粒体靶向干预措施 SS-31 对线粒体相互作用组的影响和线粒体靶向过氧化氢酶，以确定蛋白质相互作用中最重要的与年龄相关的变化网络。目标 3 测试目标 1 和 2 中确定的线粒体相互作用组的变化是否转化为老化的人体骨骼肌。老年人 SS-31 的线粒体相互作用组、功能和作用分为低表现组和高表现组，并与年轻人进行比较。这些实验的结果将产生重大影响，并有可能通过提供第一个测试来改变该领域线粒体蛋白质相互作用的破坏会导致与年龄相关的线粒体功能障碍。 1