Sequential selective enrichment of phosphopeptides and glycopeptides using amine-functionalized magnetic nanoparticles
Sequential selective enrichment of phosphopeptides and glycopeptides using amine-functionalized magnetic nanoparticles
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使用胺功能化磁性纳米粒子顺序选择性富集磷酸肽和糖肽
DOI：
10.1039/c2mb25288k
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发表时间：
2013-01-01
影响因子：
--
通讯作者：
Lu, Haojie
中科院分区：
文献类型：
--
作者：
Zhang, Ying;Wang, Huijie;Lu, Haojie
糖基化和磷酸化作为蛋白质最常见和最重要的翻译后修饰，在生物过程中起着至关重要的作用。由于磷酸/糖肽丰度较低，利用质谱仪(MS)鉴定蛋白质的磷酸化和糖基化尤其需要特异和灵敏的策略。然而，以前报道的方法大多只专注于富集磷酸肽或糖肽，而不是同时富集磷酸肽和糖肽。本研究采用一锅法制备胺功能化磁性纳米粒子，并成功地将其用于磷酸肽和糖肽的选择性富集。通过对摩尔比为10：1：1的多肽/磷酸肽/糖肽混合物的分析，证明了该方法的选择性，磷酸肽和糖肽的后富集回收率分别为88%和76%。对磷酸肽和糖肽的敏感性均在fmoL水平。此外，序列覆盖率从25.6%增加到51.8%，对应于模型蛋白质asialofetuin的102%的增加。这些新鉴定的磷酸肽或糖肽提供了额外的序列信息，这有利于蛋白质的鉴定。
As two of the most common and important post-translational modifications (PTMs) of proteins, glycosylation and phosphorylation play critical roles in biological processes. Because of the low abundance of phosphopeptides/glycopeptides, specific and sensitive strategies are especially indispensable for the identification of protein phosphorylation and glycosylation by mass spectrometry (MS). However, most of those previously reported methods only focused on enriching either phosphopeptides or glycopeptides rather than enriching both of them. In the present study, amine-functionalized magnetic nanoparticles were synthesized in a one-pot procedure and successfully used for selective enrichment of both phosphopeptides and glycopeptides. The selectivity of this method was demonstrated by analyzing the mixture of peptides/phosphopeptides/glycopeptides at molar ratio of 10 : 1 : 1; the post-enrichment recovery was 88% and 76% for phosphopeptides and glycopeptides respectively. The sensitivity was at the fmol level for both of the phosphopeptides and glycopeptides. In addition, sequence coverage was increased from 25.6% to 51.8% corresponding to a 102% increase for a model protein asialofetuin. These newly identified phosphopeptides or glycopeptides provided additional sequence information, which was beneficial to the protein identification.