Conversion of pyridylamino sugar chains to 1-amino-1-deoxy derivatives, intermediates for tagging with fluorescein and biotin.
Pyridylamino (PA) derivatives of sugar chains were converted to 1-amino-1-deoxy derivatives. PA-lactose as a model compound was reduced with hydrogen, then treated with hydrazine. The product obtained was identified as 1-amino-1-deoxylactitol by mass spectrometry and chromatography with 1-amino-1-deoxylactitol as standard. PA-N-acetylglucosamine was converted to 1-amino-1-deoxy-N-acetylglucosaminitol under the same conditions. As an application, Man alpha 1-6(Man alpha 1-3)Man alpha 1- 6(Man alpha 1-2Man alpha 1-3)-Man beta 1-4GlcNAc beta 1-4GlcNAc-PA was converted to the 1-amino-1-deoxy derivative, which was further derivatized with fluorescein isothiocyanate or biotin sulfo-N-hydroxy-succinimide ester. Binding of these derivatives to concanavalin A dot-blotted on a nitrocellulose membrane was confirmed by fluorescence and by streptavidin-peroxidase conjugate. This conversion allowed replacement of the PA-group in PA-sugar chains which can be easily purified from glycoconjugates.
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Department of Chemistry, Osaka University College of Science.
D000606:Amino Sugars / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002852:Chromatography, Ion Exchange; D019793:Fluorescein; D005452:Fluoresceins / Q000737:chemistry*; D006834:Hydrazines / Q000737:chemistry; D006859:Hydrogen / Q000737:chemistry; D007785:Lactose / Q000737:chemistry*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction
D000606:Amino Sugars; D005452:Fluoresceins; D006834:Hydrazines; C029424:hydrazine; D001710:Biotin; D006859:Hydrogen; D007785:Lactose; D019793:Fluorescein