Separation of Galf beta 1 -> XGlcNAc and Galp beta 1 -> XGlcNAc (X=3, 4, and 8) as the alditols by high-pH anion-exchange chromatography and thin-layer chromatography: Characterization of mucins from Trypanosoma cruzi

The O-linked N-acetylglucosamine oligosaccharides in the mucins of Trypanos oma cruzi may contain galactofuranose or galactopyranose, depending on the strain, one of the components being the disaccharide, Since galactofuranose is a site for antibody recognition, it is desirable to have a sensitive me thod for the detection of the galactofuranosyl structures. In this paper, w e present procedures for the separation of Galf beta 1-->XGlcNAc and Galp b eta 1-->XGlcNAc (X = 3, 4, and 6) as the corresponding alditols by high-pH anion-exchange chromatography with pulse amperometric detection, AU the iso meric disaccharides could be resolved on a CarboPac PA-10 column, the galac tofuranose-containing disaccharides being more retained in the column, GlcN Acol and Galf beta 1-->4(Galp beta 1-->6)GlcNAcol could be analyzed in the same run, The compounds could also be separated by thin-layer chromatograph y on silica gel 60, a convenient method for analysis of the radio-labeled a lditols obtained by reductive beta-elimination in the presence of (NaBH4)-H -3. Both methods were applied for the analysis of the O-linked sugars in th e mucins of T. cruczi CL 14 and revealed that they contained only N-acetylg lucosamine and the disaccharide Galp beta 1--> 4GlcNAc. (C) 2000 Academic P ress.