Tandem Mass Spectrometric Characterization of Fetuin Sialylated Glycopeptides Enriched by TiO2 Microcolumn

Sialylation of glycoproteins is vital for the function or physicochemical properties of a protein. It becomes more and more important to develop approaches that can be used to efficiently isolate and identify sialylated glycopeptides or glycoproteins for monitoring changes in glycoproteome. In the present study, we analyze intact structures of the enriched sialylated glycopeptides of bovine fetuin by matrix-assisted laser desorption/ionization-tandem mass spectrometry (MALDI-MS/MS), without any chemical derivation. The experimental data show that the optimal loading buffer for TiO2 as matrix is 80% acetonitrile/2% TFA (trifluoroacetic acid)/100 mg/mL DHB (2,5-dihydroxybenzoic acid) which is also compatible with MALDI-mass spectrometric analysis. This study indicates that the improved enrichment approach combined with MALDI-MS/MS may be a powerful tool to analyze intact structures and components of the sialylated glycopeptides from complex peptide mixture.