Separation of Galfbeta1-->XGlcNAc and Galpbeta1-->XGlcNAc (X = 3, 4, and 6) 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 Trypanosoma 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 method for the detection of the galactofuranosyl structures. In this paper, we present procedures for the separation of Galfbeta1-->XGlcNAc and Galpbeta1-->XGlcNAc (X = 3, 4, and 6) as the corresponding alditols by high-pH anion-exchange chromatography with pulse amperometric detection. All the isomeric disaccharides could be resolved on a CarboPac PA-10 column, the galactofuranose-containing disaccharides being more retained in the column. GlcNAcol and Galfbeta1-->4(Galpbeta1-->6)GlcNAcol could be analyzed in the same run. The compounds could also be separated by thin-layer chromatography on silica gel 60, a convenient method for analysis of the radiolabeled alditols obtained by reductive beta-elimination in the presence of NaB(3)H(4). Both methods were applied for the analysis of the O-linked sugars in the mucins of T. cruzi CL 14 and revealed that they contained only N-acetylglucosamine and the disaccharide Galpbeta1-->4GlcNAc.
Salto
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ML
CIHIDECAR, Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Pabellón II, Buenos Aires, 1428, Argentina.
Gallo-Rodriguez
C
C
Lima
C
C
de Lederkremer
R M
RM
D000818:Animals; D002240:Carbohydrate Sequence; D002852:Chromatography, Ion Exchange / Q000379:methods*; D002855:Chromatography, Thin Layer / Q000379:methods*; D004187:Disaccharides / Q000737:chemistry / Q000302:isolation & purification*; D005069:Evaluation Studies as Topic; D006863:Hydrogen-Ion Concentration; D009077:Mucins / Q000737:chemistry* / Q000302:isolation & purification; D013402:Sugar Alcohols / Q000737:chemistry / Q000302:isolation & purification; D014349:Trypanosoma cruzi / Q000737:chemistry*
D004187:Disaccharides; D009077:Mucins; D013402:Sugar Alcohols