Mapping O-Glycosylation Sites of Secreted Proteins by Combining Lectin Chromatography, Enzymatic Deglycosylation and Multistage Mass Spectrometry

摘要

Two step Jacalin enrichment on both protein and peptide level gives a good enrichment of O-glycosylated peptides. Partial enzymatic deglycosylation simpliifes analysis of both O- and N-glycosylated peptides. The carbohydrate attachment site can be determined using the residual glycan. Sequential MS2/MS3 allows peptide identitcation. ECD is possible on an lc-timescale and gives invaluable information on the glycan attachment sites. Drawbacks are relatively low sensitivity and sparsely distributed spectra of proline rich peptides.