SH2 Superbinder Modified Monolithic Capillary Column for the Sensitive Analysis of Protein Tyrosine Phosphorylation

• DOI: 10.1021/acs.jproteome.7b00546
• Publication Date: 2017-10-30T15:01:48Z
• Authors: Yating Yao; Yangyang Bian; Mingming Dong; Yan Wang; Jiawen Lv; Lianfang Chen; Hongwei Wang; Jiawei Mao; Jing Dong; Mingliang Ye
• Keywords: Microreactor; Phosphopeptide; Phosphoproteomics

In this study, we present a method to specifically capture phosphotyrosine (pTyr) peptides from minute amount of sample for the sensitive analysis protein tyrosine phosphorylation. We immobilized SH2 superbinder on monolithic capillary column construct microreactor enrich pTyr peptides. It was found that synthetic peptide could be enriched by mixture prepared spiking into tryptic digests α-casein and β-casein with molar ratios 1:1000:1000. The further applied pervanadate-treated HeLa cell phosphoproteomics analysis, which resulted in identification 796 unique sites. contrast, conventional superbinder-based identified 41 sites same sample, only 5.2% number achieved microreactor. Finally, also analyze Shc1 complex, an immunopurified 15 Together, technique is best fitted will have broad application fields where limited available.