基于液相色谱-串联质谱技术的磷酸化蛋白质组学分析

Phosphoproteomic Analysis Based on Liquid Chromatography-Tandem Mass Spectrometry

本文运用稳定同位素双甲基化标记技术、固定相金属离子螯合层析（IMAC）技术并结合液相色谱-串联质谱法研究了仙台病毒感染引起的宿主细胞磷酸化蛋白质组变化。实验发现定量的4 289个磷酸化肽段有~20%的蛋白质磷酸化位点发生了显著变化;通路富集分析表明哺乳动物雷帕毒素靶蛋白（mTOR）通路和剪接体（Spliceosome）通路可能参与宿主细胞对病毒的应答;通过对显著变化磷酸化肽段进行基序分析,发现了9个代表性的磷酸化修饰位点基序。本研究方法对于深入解析抗病毒天然免疫信号通路提供了新线索。

In this study,we utilized stable isotope dimethylated labeling,immsbilized metal ion affinity chromatography（IMAC）based phosphopeptide enrichment,and liquid chromatography-tandem mass spectrometry（LC-MS/MS）to quantitate the host phosphoproteome changes after SeV infection of host cells.We quantified 4 289 phosphopeptides,among which,~20% phosphorylation sites were found as significantly changed upon viral infection.Pathway enrichment analysis showed that mTOR and spliceosome pathways may be involved in antiviral response.Phosphorylation motif analysis revealed 9representative motifs were present in the phosphorylation sites that have been found.This study provides new insights into antiviral innate immune signaling pathway.