Improved sequence tag generation method for peptide identification in tandem mass spectrometry.

Improved sequence tag generation method for peptide identification in tandem mass spectrometry.
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DOI:
10.1021/pr800400q
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发表时间:
2008-10
影响因子:
4.4
通讯作者:
Nesvizhskii, Alexey I.
Nesvizhskii, Alexey I.
中科院分区:
生物学2区
文献类型:
--
作者:
Cao, Xia;Nesvizhskii, Alexey I.

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基于序列标签的多肽鉴定方法是传统数据库搜索方法的一种有前途的替代方法。然而,更全面的分析,优化,并与已建立的方法进行比较,这些方法可以在蛋白质组学社区获得广泛使用之前是必要的。使用InsPecT开放源代码库(坦纳等人,Anal Chem.2005,77:4626-39),我们提出了一种改进的序列标签生成方法,其直接并入存在于许多更高电荷状态的串联质谱中的多电荷碎片离子峰。我们还研究了在不同设置下使用五种不同类型的质谱仪生成的控制数据集以及使用复杂的磷酸肽富集样品的序列标记的性能。我们还证明,使用半参数方法结合前体离子质量测量的准确性的InsPecT搜索分数的额外建模提供了区分正确和不正确的肽鉴定的能力的额外改进。通过与常用的SEQUEST/PeptideProphet方法进行比较,证明了基于序列标签的肽鉴定方法的总体上级性能。
The sequence tag-based peptide identification methods are a promising alternative to the traditional database search approach. However, a more comprehensive analysis, optimization, and comparison with established methods are necessary before these methods can gain widespread use in the proteomics community. Using the InsPecT open source code base (Tanner et al., Anal Chem. 2005, 77:4626–39), we present an improved sequence tag generation method that directly incorporates multi-charged fragment ion peaks present in many tandem mass spectra of higher charge states. We also investigate the performance of sequence tagging under different settings using control datasets generated on five different types of mass spectrometers, as well as using a complex phosphopeptide-enriched sample. We also demonstrate that additional modeling of InsPecT search scores using a semi-parametric approach incorporating the accuracy of the precursor ion mass measurement provides additional improvement in the ability to discriminate between correct and incorrect peptide identifications. The overall superior performance of the sequence tag-based peptide identification method is demonstrated by comparison with a commonly used SEQUEST/PeptideProphet approach.
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