Protein turnover: Measurement of proteome dynamics by whole animal metabolic labelling with stable isotope labelled amino acids
Protein turnover: Measurement of proteome dynamics by whole animal metabolic labelling with stable isotope labelled amino acids
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DOI：
10.1002/pmic.201100556
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发表时间：
2012-04-01
期刊：
影响因子：
3.4
通讯作者：
Beynon, Robert J.
中科院分区：
文献类型：
--
作者：
Claydon, Amy J.;Thom, Michael D.;Beynon, Robert J.
完整动物组织中蛋白质周转的测量是通过全动物动态标记研究获得的，需要通过饮食给予前体标记。很难获得饮食中前体氨基酸的完整标记，如果使用部分标记，则计算每种蛋白质的周转率需要了解前体相对同位素丰度（RIA）。我们描述了一种方法，以动态标记的蛋白质在小鼠与商业饮食补充了一个纯的，氘代必需氨基酸。同位素标记模式可用于恢复前体RIA，尿分泌蛋白采样可无创监测肝脏前体RIA的发展。单个蛋白质的标记轨迹的时间序列分析允许准确测定降解的一阶速率常数。在多个蛋白质上获得该参数允许细胞蛋白质的周转谱和不同组织的比较。肌肉蛋白质降解的中位速率远低于肝脏或肾脏，心脏处于中间位置。
The measurement of protein turnover in tissues of intact animals is obtained by whole animal dynamic labelling studies, requiring dietary administration of precursor label. It is difficult to obtain full labelling of precursor amino acids in the diet and if partial labelling is used, calculation of the rate of turnover of each protein requires knowledge of the precursor relative isotope abundance (RIA). We describe an approach to dynamic labelling of proteins in the mouse with a commercial diet supplemented with a pure, deuterated essential amino acid. The pattern of isotopomer labelling can be used to recover the precursor RIA, and sampling of urinary secreted proteins can monitor the development of liver precursor RIA non-invasively. Time-series analysis of the labelling trajectories for individual proteins allows accurate determination of the first order rate constant for degradation. The acquisition of this parameter over multiple proteins permits turnover profiling of cellular proteins and comparisons of different tissues. The median rate of degradation of muscle protein is considerably lower than liver or kidney, with heart occupying an intermediate position.