Separation of Galfβ1→XGlcNAc and Galpβ1→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
✍ Scribed by Marı́a Laura Salto; Carola Gallo-Rodriguez; Carlos Lima; Rosa M. de Lederkremer
- Publisher
- Elsevier Science
- Year
- 2000
- Tongue
- English
- Weight
- 108 KB
- Volume
- 279
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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 Galf13 XGlcNAc and Galp13XGlcNAc (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 Galf134(Galp136)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 -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 Galp13 4GlcNAc.