Broad range of missense error frequencies in cellular proteins
Broad range of missense error frequencies in cellular proteins
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DOI：
10.1093/nar/gky1319
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发表时间：
2019-04-08
影响因子：
14.9
通讯作者：
Rodnina, Marina, V
中科院分区：
文献类型：
--
作者：
Garofalo, Raffaella;Wohlgemuth, Ingo;Rodnina, Marina, V
评估基因表达的保真度对于了解细胞内环境的稳定至关重要。在此，我们提出了一种高灵敏度的系统定量稀有氨基酸取代(QRAS)的方法，通过对含有错义氨基酸取代的多肽进行色谱层析富集后，用靶向质谱仪进行绝对定量。通过分析一个模型蛋白质EF-Tu中的近源氨基酸和非同源氨基酸的掺入情况，我们发现大多数错义错误太少了，无法用传统的方法，如DDA来检测，估计在
Assessment of the fidelity of gene expression is crucial to understand cell homeostasis. Here we present a highly sensitive method for the systematic Quantification of Rare Amino acid Substitutions (QRAS) using absolute quantification by targeted mass spectrometry after chromatographic enrichment of peptides with missense amino acid substitutions. By analyzing incorporation of near- and non-cognate amino acids in a model protein EF-Tu, we show that most of missense errors are too rare to detect by conventional methods, such as DDA, and are estimated to be between