Age induced interactome remodeling in muscle mitochondria

A novel quantitative crosslinking mass spectrometry technique (qXL-MS) was applied to elucidate interactome changes in aged murine skeletal muscle mitochondria that contribute to age-related mitochondrial functional decline. Reported here are results from initial investigations of murine muscle mitochondrial interactomes that enable identification of statistically significant changes associated with aging. Newly developed isobaric quantitative protein interaction reporter (iqPIR) technologies15 enabled reproducible detection of age-related mitochondrial interactome changes. Muscle mitochondria from young and old mice were isolated, cross-linked with iqPIR molecules, young and old mitochondrial samples were paired, process and analyzed to quantify age-related mitochondrial interactome changes. In parallel, mitochondria were also subjected to additional measurements of mitochondrial protein yield, functional measurements including oxygen consumption rates on Complex I and Complex II substrates, and citrate synthase activity.

本研究采用一种新型定量交联质谱技术（quantitative crosslinking mass spectrometry，qXL-MS），以阐明衰老小鼠骨骼肌线粒体中参与年龄相关线粒体功能衰退的相互作用组变化。本文报道了小鼠肌肉线粒体相互作用组的初步研究成果，该成果可用于识别与衰老相关的具有统计学显著性的变化。新开发的同量异位素定量蛋白质相互作用报告因子（isobaric quantitative protein interaction reporter，iqPIR）技术[15] 可重复性检测到与衰老相关的线粒体相互作用组变化。研究人员分离了年轻与老年小鼠的肌肉线粒体，使用iqPIR分子进行交联处理，随后将年轻与老年线粒体样品进行配对、处理并分析，以此量化与衰老相关的线粒体相互作用组变化。与此同时，研究人员还对分离得到的线粒体开展了额外检测：包括线粒体蛋白产量测定、以复合物I（Complex I）和复合物II（Complex II）为底物的耗氧率功能检测，以及柠檬酸合酶活性测定。