Integrated Methodology for Proteomics Analysis of Glycoproteins

摘要

Glycosylation is one of the most common post-translational modification of proteins; however robust MS-based strategies for glycoproteome studies are still lacking. Challenges in glycoproteomics include: localization of the glycosylation site on the peptide backbone, the determination of the site occupancy, and ideally the elucidation of the structure of the glycans. In this context, a global workflow combining the enrichment for glycopeptides and their characterization using LC-MS platforms was developed. Partial enzymatic deglycosylation leaving an acetylglucosamine (GlcNAc) residue attached to the peptide facilitates detection of the glycopeptides using LC-MS, and enables precise identification by electron transfer dissociation (ETD) sequencing.