Tatsuta Takashi, Langer Thomas
Institut für Genetik, Universität zu Köln, Germany.
Methods Mol Biol. 2007;372:343-60. doi: 10.1007/978-1-59745-365-3_25.
Mitochondria are dynamic organelles with activities that adjust to altering physiological conditions and variable metabolic demands. A conserved proteolytic system present within the organelle exerts essential functions during the biogenesis of mitochondria and ensures the maintenance of organellar activities under varying conditions. Proteases dependent on adenosine triphosphate, in concert with oligopeptidases, degrade nonassembled or damaged proteins in various subcompartments of mitochondria, preventing their accumulation and possibly deleterious effects on mitochondrial functions. Although an increasing number of mitochondrial peptidases are characterized and functionally linked to diverse cellular processes, only limited information is available on the stability of the mitochondrial proteome and the turnover rates of individual proteins. We describe experimental approaches in the yeast Saccharomyces cerevisiae and in mice, allowing analysis of the proteolytic breakdown of mitochondrial proteins individually or on a proteomewide scale.
线粒体是动态细胞器，其活动会根据生理条件的变化和不同的代谢需求进行调整。细胞器内存在的一个保守的蛋白水解系统在 mitochondria 的生物发生过程中发挥着重要功能，并确保在不同条件下细胞器活动的维持。依赖三磷酸腺苷的蛋白酶与寡肽酶协同作用，在线粒体的各个亚区室中降解未组装或受损的蛋白质，防止它们的积累以及对线粒体功能可能产生的有害影响。尽管越来越多的线粒体肽酶被鉴定出来并在功能上与多种细胞过程相关联，但关于线粒体蛋白质组的稳定性和单个蛋白质的周转速率的信息仍然有限。我们描述了在酿酒酵母和小鼠中的实验方法，这些方法允许单独或在蛋白质组范围内分析线粒体蛋白质的蛋白水解降解情况。
