期刊
NUCLEIC ACIDS RESEARCH
卷 47, 期 6, 页码 2932-2945出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gky1319
关键词
-
资金
- German Science Foundation (Deutsche Forschungsgemeinschaft) via Research Unit FOR [1805]
- Max Planck Society
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 <10(-7)-10(-5) by QRAS. We also observe error hotspots of up to 10(-3) for some types of mismatches, including the G-U mismatch. The error frequency depends on the expression level of EF-Tu and, surprisingly, the amino acid position in the protein. QRAS is not restricted to any particular miscoding event, organism, strain or model protein and is a reliable tool to analyze very rare proteogenomic events.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据