Metal ion-decorated ZIC-cHILIC StageTip Enables Simultaneous Profiling of N-Glycoproteome and Phosphoproteome in TKI resistant NSCLC cells

Cell membrane proteins are densely decorated with surface glycosylation and intracellular phosphorylation whose interplay determines the cell-cell communication and signaling cascades. However, their concomitant characterization remains extreme challenges due to complexity of glycan structures, low abundance of glycopeptides (particularly sialylation), labile, dynamic nature and low detectability of both phosphopeptides and glycopeptides in mass spectrometry. In addition, the interplay and dynamic change between glycosylation and phosphorylation in tyrosine kinase inhibitor (TKI)-resistant non-small cell lung cancer (NSCLC) cells is still unclear. In this study, we introduce a streamlined metal ion-decorated ZIC-cHILIC strategy, featuring a simple pH control, to allow simultaneous enrichment and stepwise separation of intact (sialo-)glycopeptides and phosphopeptides.