O-linked glycans are characterized as smaller but highly diverse branched carbohydrates molecules rather than N-linked glycans and usually attach to proteins at any supposed field, therefore, it lead to a complexity of analysis and is difficult to pinpoint these exact locations of O-glycosylation.
We can either carry out a summary analysis of all glycosylation sites or analysis of site-specific glycosylation.
The general steps for O-glycosylation site occupation analysis inclueds proteolytical digestion of glycoprotein, a precipitation step, glycopeptide enrichment and fractionation via hydrophilic interaction liquid chromatography. Enriched O-glycopeptide fractions were analyzed by mass spectrometric analysis using reversed-phase liquid chromatography coupled ion trap mass spectrometer.
Figure 1. Workflow for the analysis of O-glycosylation site occupancy
Other label free approaches based on the signal intensity of glycopeptide/peptide ions for analysis of O-glycosylation site occupation is as well available.
As one of the leading companies in the omics field with over years of experience in omics study, Creative Proteomics provides glycomics analysis service customized to your needs. Contact us to discuss your project.
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