RAPID DETECTION OF THE ALTERNATIVE N-GLYCOSYLATION PATHWAY USING HIGHPH ANION-EXCHANGE CHROMATOGRAPHY

An alternative N-glycosylation pathway using Glc(1-3)Man(5)GlcNAc(2) a s a donor to be transferred to a protein acceptor is found either in M an-P-Dol synthase deficient cells or in wild type CHO cells grown in e nergy deprivation conditions. Discrimination between oligomannosides o f this alternative pathway and oligomannosides of the major one contai ning the same number of sugar residues Man(6-8)GlcNAc(2) required stru ctural studies. Taking advantage of the specific chromatographic behav iour of glucosylated oligomannosides, in pellicular high pH anion exch ange chromatography, we developed a one-step method for the identifica tion of the alternative N-glycosylation pathway compounds differing fr om those of the major one.