Abstract
Hyaluronidase from the venom of the honeybee (Apis mellifera) has been purified by gelpermeation and cation exchange chromatography. Its asparagine-linked carbohydrate chains were released from tryptic glycopeptides with N-glycosidase A and reductively aminated with 2-aminopyridine. Separation of the fluorescent derivatives by size-fractionation and reversed-phase HPLC afforded eighteen fractions which were analysed by two-dimensional HPLC mapping combined with exoglycosidase digestions. The bulk of the N-linked glycans of hyaluronidase consisted of small oligosaccharides (Man1–3GlcNAc2), most of which were either α1,3-monofucosylated or α1,3-(α1,6-)difucosylated at the innermost GlcNAc residue. High-mannose type structures constituted the minor fractions, together making up about 5% of the oligosaccharide pool from hyaluronidase. Four fractions, making up 8% of the N-linked glycans, contained the terminal trisaccharide GalNAcβ1-4[Fucα1-3]GlcNAcβ1- in β1,2-linkage to the core α1,3-mannosyl residue. No evidence for the presence of O-glycans or sialic acids could be found.
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Abbreviations
- GalNAc:
-
N-acetylgalactosamine
- GlcNAc:
-
N-acetylglucosamine
- PA:
-
pyridylamino
- PLA:
-
phospholipase A2
- 2D-HPLC:
-
two-dimensional HPLC
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Kubelka, V., Altmann, F. & März, L. The asparagine-linked carbohydrate of honeybee venom hyaluronidase. Glycoconjugate J 12, 77–83 (1995). https://doi.org/10.1007/BF00731872
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DOI: https://doi.org/10.1007/BF00731872