Impact of Amidination on Peptide Fragmentation and Identification in Shotgun Proteomics.

Impact of Amidination on Peptide Fragmentation and Identification in Shotgun Proteomics.
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DOI:
10.1021/acs.jproteome.6b00468
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发表时间:
2016-10-07
影响因子:
4.4
通讯作者:
Reilly JP
Reilly JP
中科院分区:
生物学2区
文献类型:
--
作者:
Li S;Dabir A;Misal SA;Tang H;Radivojac P;Reilly JP

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肽脒标记使用S-甲基硫代乙酰亚胺(SMTA)的研究,试图增加的数量和类型的肽,可以检测到在自下而上的蛋白质组学实验。该衍生方法影响赖氨酸残基的碱性,并且在此显示显著影响肽片段化和肽可检测性的特异性。SMTA标记肽的独特和高度可重复的片段化特性,如形成b1碎片离子的强烈倾向,可进一步用于修改肽-光谱对的评分并改善肽鉴定。为此,我们开发了一种监督后处理算法,利用这些特点的肽标记SMTA。我们的实验表明,虽然鉴定的总数是相似的,但SMTA修饰使得能够检测到16-26%的肽,这是以前在没有SMTA标记的可比CID/HCD串联质谱实验中未观察到的。
Peptide amidination labeling using S-methyl thioacetimidate (SMTA) is investigated in an attempt to increase the number and types of peptides that can be detected in a bottom-up proteomics experiment. This derivatization method affects the basicity of lysine residues, and is shown here to significantly impact the idiosyncracies of peptide fragmentation and peptide detectability. The unique and highly reproducible fragmentation properties of SMTA-labeled peptides, such as the strong propensity for forming b1 fragment ions, can be further exploited to modify the scoring of peptide-spectrum pairs and improve peptide identification. To this end, we have developed a supervised post-processing algorithm to exploit these characteristics of peptides labeled by SMTA. Our experiments show that, although the overall number of identifications are similar, the SMTA modification enabled the detection of 16-26% peptides not previously observed in comparable CID/HCD tandem mass spectrometry experiments without SMTA labeling.
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