Title : Proteome-wide characterization of N-glycosylation events by diagonal chromatography
Abstract :
- A procedure to map N-glycosylation sites is presented here
- It can be applied to purified proteins as well as to highly complex mixtures
- The method exploits deglycosylation by PNGase F in a diagonal, reverse-phase chromatographic setup
- When applied to 10 microL of mouse serum, affinity-depleted for its three most abundant components, 117 known or predicted sites were mapped in addition to 10 novel sites
- Several sites were detected on soluble membrane or receptor components
- Our method furthermore senses the nature of glycan structures and can detect differential glycosylation on a given site
- These properties--high sensitivity and dependence on glycan imprinting--can be exploited for glycan-biomarker analysis