Impact of ion trap tandem mass spectra variability on the identification of peptides
Impact of ion trap tandem mass spectra variability on the identification of peptides
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DOI：
10.1021/ac0348219
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发表时间：
2004-05-15
影响因子：
7.4
通讯作者：
Yates, JR
中科院分区：
文献类型：
--
作者：
Venable, JD;Yates, JR
基于串联质谱学和数据库搜索算法的多肽鉴定已成为蛋白质组学的核心技术之一。这项技术的核心是能够重复获得用于数据库查询的高质量串联质谱图。串联质谱图的可变性通常被认为是最小的，并且肽的鉴定通常基于单个串联质谱图。在这篇文章中，我们用几种数据库搜索算法表征了从重复串联质谱学得到的分数的方差，并展示了光谱变异性对正确识别多肽的影响。我们表明，与串联质量谱集合相关的方差可能是相当大的，从而导致搜索算法得分中的相当大的误差(类似于5-25%的RSD)，并最终导致不正确的分配。讨论了处理策略，以最大限度地减少串联质谱可变性对肽鉴定的影响。
Peptide identification based on tandem mass spectrometry and database searching algorithms has become one of the central technologies in proteomics. At the heart of this technology is the ability to reproducibly acquire high-quality tandem mass spectra for database interrogation. The variability in tandem mass spectra generation is often assumed to be minimal, and peptide identifications are typically based on a single tandem mass spectrum. In this paper, we characterize the variance of scores derived from replicate tandem mass spectra using several database search algorithms and demonstrate the effects of spectral variability on the correct identification of peptides. We show that the variance associated with the collection of tandem mass spectra can be substantial leading to sizable errors in search algorithm scores (similar to5-25% RSD) and ultimately incorrect assignments. Processing strategies are discussed to minimize the impact of tandem mass spectra variability on peptide identification.