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首页> 外文期刊>Journal of mass spectrometry: JMS >Using spectral matching to interpret LC-MS/MS data during RNA modification mapping
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Using spectral matching to interpret LC-MS/MS data during RNA modification mapping

机译：使用光谱匹配在RNA修改映射期间解释LC-MS / MS数据

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摘要

While a number of approaches have been developed to analyze liquid chromatography tandem mass spectrometry (LC-MS/MS) data obtained from modified oligonucleotides, the majority of these methods require analyzing every MS/MS spectrum de novo to sequence the oligonucleotide and place the modification. Spectral matching is an alternative approach for analyzing MS/MS data that is based on creating a library of annotated MS/MS spectra against which individual MS/MS data can be searched. Here, we have adapted the existing NIST spectral matching software to enable its use in the interpretation of MS/MS data obtained from modified oligonucleotides. In particular, we demonstrate the utility of this approach to identify specific post-transcriptionally modified nucleosides in particular transfer RNAs (tRNAs) obtained through a conventional RNA modification mapping experimental protocol. Spectral matching was found to be an efficient approach for screening for known modified tRNAs by using the experimental data as the library and previously annotated RNase T1 digestion products of tRNAs as the reference spectra. The utility of spectral matching for rapid analysis of multiple LC-MS/MS analyses was demonstrated by screening mutant strains of Streptococcus mutans to identify the enzyme(s) responsible for synthesizing the tRNA position 37 modification threonylcarbamoyladenosine (t(6)A).
机译：虽然已经开发了许多方法来分析从改性寡核苷酸获得的液相色谱串联质谱（LC-MS / MS）数据，但这些方法中的大多数需要分析每个MS / MS光谱DE Novo序列寡核苷酸并放置改性。光谱匹配是用于分析基于创建注释的MS / MS谱图集的MS / MS数据的替代方法，该数据可以搜索各个MS / MS数据。在这里，我们已经改编了现有的NIST光谱匹配软件，以使其在从改性寡核苷酸获得的MS / MS数据的解释中使用。特别是，我们证明了这种方法鉴定了通过常规RNA修改映射实验方案获得的特定转移RNA（TRNA）的特定转录后修饰的核苷的效用。发现光谱匹配是通过使用实验数据作为文库的实验数据和TRNA的先前注释的RNase T1消化产物作为参考光谱来筛选已知改性TRNA的有效方法。通过筛选链球菌的突变体菌株来证明对多LC-MS / MS分析进行快速分析的效用，以鉴定负责合成TRNA位置37改性苏氨基氨基甲酰胺（T（6）A）的酶。

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