Recent advances in demystifying O‐glycosylation in health and disease

O‐Glycosylation is one of the most common protein post‐translational modifications (PTM) and plays an essential role in the pathophysiology of diseases. However, the complexity of O‐glycosylation and the lack of specific enzymes for the processing of O‐glycans and their O‐glycopeptides make O‐glycosylation analysis challenging. Recently, research on O‐glycosylation has received attention owing to technological innovation and emerging O‐glycoproteases. Several serine/threonine endoproteases have been found to specifically cleave O‐glycosylated serine or threonine, allowing for the systematic analysis of O‐glycoproteins. In this review, we first assessed the field of protein O‐glycosylation over the past decade and used bibliometric analysis to identify keywords and emerging trends. We then summarized recent advances in O‐glycosylation, covering several aspects: O‐glycan release, site‐specific elucidation of intact O‐glycopeptides, identification of O‐glycosites, characterization of different O‐glycoproteases, mass spectrometry (MS) fragmentation methods for site‐specific O‐glycosylation assignment, and O‐glycosylation data analysis. Finally, the role of O‐glycosylation in health and disease was discussed.

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