Lombardi A, Silvestri E, Cioffi F, Senese R, Lanni A, Goglia F, de Lange P, Moreno M
Dipartimento delle Scienze Biologiche, Sezione Fisiologia, Università degli Studi di Napoli Federico II, Via Mezzocannone 8, Naples, Italy.
J Proteomics. 2009 May 2;72(4):708-21. doi: 10.1016/j.jprot.2009.02.007. Epub 2009 Mar 5.
We defined the transcriptomic and proteomic profiles of rat ageing skeletal muscle using a combined cDNA array, 2D- and Blue native-PAGE approach. This was allowed to obtain an overview of the interrelated events leading to the transcriptome/proteome/mitoproteome changes likely to underlie the structural/metabolic features of aged skeletal muscle. The main differences were found in genes/proteins related to energy metabolism, mitochondrial pathways, myofibrillar filaments, and detoxification. Concerning the abundance of mitochondrial OXPHOS complexes as well as their supramolecular organization and activity, mitochondria from old rats, when compared with those from young rats, contained significantly lower amounts of complex I (NADH:ubiquinone oxidoreductase), V (FoF1-ATP synthase), and III (ubiquinol:cytochrome c oxidoreductase). The same mitochondria contained a significantly larger amount of complex II (succinate:ubiquinone oxidoreductase), but an unchanged amount of complex IV (cytochrome c oxidase, COX). When comparing the supercomplex profiles between young and old muscle mitochondria, the densitometric analysis revealed that lighter supercomplexes were significantly reduced in older mitochondria, and that in the older group the major supercomplex bands were those representing heavier supercomplexes, likely suggesting a compensatory mechanism that, in ageing muscle, is functionally directed towards substrate channeling and catalytic enhancement advantaging the respirosome.
我们使用组合cDNA阵列、二维和蓝色原胶聚丙烯酰胺凝胶电泳方法定义了大鼠衰老骨骼肌的转录组和蛋白质组图谱。这使得我们能够概述导致转录组/蛋白质组/线粒体蛋白质组变化的相关事件,这些变化可能是衰老骨骼肌结构/代谢特征的基础。主要差异存在于与能量代谢、线粒体途径、肌原纤维丝和解毒相关的基因/蛋白质中。关于线粒体氧化磷酸化复合物的丰度及其超分子组织和活性,与年轻大鼠的线粒体相比,老年大鼠的线粒体中复合物I(NADH:泛醌氧化还原酶)、V(FoF1 - ATP合酶)和III(泛醇:细胞色素c氧化还原酶)的含量显著降低。相同的线粒体中复合物II(琥珀酸:泛醌氧化还原酶)的含量显著增加,但复合物IV(细胞色素c氧化酶,COX)的含量不变。在比较年轻和老年肌肉线粒体的超复合物图谱时,光密度分析显示,较轻的超复合物在老年线粒体中显著减少,并且在老年组中,主要的超复合物条带是代表较重超复合物的条带,这可能表明一种补偿机制,在衰老肌肉中,该机制在功能上导向底物通道化和催化增强,有利于呼吸体。