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Phosphoproteomic Analysis Based on Liquid Chromatography-Tandem Mass Spectrometry

ZHU Xiu-juan;LI Hao-yu;GUO Lin;ZHAO Xiao-lu;State Key Laboratory of Virology,College of Life Sciences,Wuhan University;  
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.
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