Aging Mitochondrial Interactome

衰老线粒体相互作用组

• 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
• 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

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学、恢复力的丧失和衰老中起着核心作用。 老年人生活质量下降。随着年龄的增长，我们的线粒体发生了变化， 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 稳态，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 组织功能障碍线粒体是动态的细胞器，不断适应不断变化的细胞需求 通过改变蛋白质的组装和相互作用来改变它们的功能。尽管重要性显而易见， 已知线粒体蛋白质相互作用（相互作用组）中与年龄相关的变化是如何导致 功能随着年龄的增长。为了解决这一基本问题，我们建议采用最先进的定量方法， 化学交联与质谱（qXL-MS）策略，以量化线粒体中的变化 与年龄相互作用。通过将这种创新的qXL-MS方法与线粒体的详细测定相结合， 代谢和干预措施，我们已经开发了在过去几年来操纵线粒体在体内 在体外，我们独特地定位于识别线粒体相互作用组的分子水平变化， 与年龄相关的线粒体功能障碍我们的初步数据表明， 参与ADP转运、ATP合成和电子传递底物供应的相互作用网络 老年人心脏和骨骼肌中的神经系统。这些变化与先前证明的 ATP生成减少，对ADP的敏感性降低。此外，我们已经证明， 靶向干预（SS-31），逆转心脏和骨骼肌中的线粒体功能障碍， 与许多相同的蛋白质复合物相互作用，我们的相互作用组学研究表明，这些蛋白质复合物在衰老中被破坏， 包括ANT和电子传递系统的络合物IV和V。我们的整体假设在这个实验中得到了验证 有人提出，线粒体相互作用组随年龄的变化是ATP产生减少的基础， 老年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， 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的变化， 在体内和体外小鼠心脏和骨骼肌中由促氧化剂处理诱导的那些。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年龄相关变化 网络.目标3测试了目标1和目标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 被分为低绩效和高绩效的群体与年轻人进行比较。这些实验的结果 将产生重大影响，并有可能通过提供如何改变该领域的第一个测试， 线粒体蛋白相互作用的破坏导致与年龄相关的线粒体功能障碍。 1