Out with the old, in with the new? Comparing methods for measuring protein degradation.
Out with the old, in with the new? Comparing methods for measuring protein degradation.
复制标题
DOI：
10.1042/cbi20110055
复制
发表时间：
2011-05
影响因子：
3.9
通讯作者：
Nicchitta CV
中科院分区：
文献类型：
--
作者：
Yewdell JW;Lacsina JR;Rechsteiner MC;Nicchitta CV
蛋白质降解是控制细胞中蛋白质丰度的关键变量。在这里，我们将测量蛋白质降解率的经典方法与一种新的基于GFP报告文库的方法进行了比较，该方法通过流式细胞术表征了数千种单个蛋白质的降解。虽然没有一种方法是完美的，但我们得出结论，嵌合基因报告方法应该谨慎应用，主要是因为GFP（或其他标签）干扰细胞器靶向或将嵌合蛋白掺入大分子组装中，导致虚假的高降解率。
Protein degradation is a key variable in controlling protein abundance in cells. Here, we compare classical methods to measure protein degradation rates with a novel GFP reporter library based method that characterizes degradation of thousands of individual proteins by flow cytometry. While no method is perfect, we conclude that chimeric gene reporter approaches should be applied cautiously due principally to GFP (or other tag) interference with organelle targeting or incorporation of chimeric proteins into macromolecular assemblies that results in spuriously high degradation rates.