Glycoconjugate Journal

, Volume 12, Issue 1, pp 77–83 | Cite as

The asparagine-linked carbohydrate of honeybee venom hyaluronidase

  • Viktoria Kubelka
  • Friedrich Altmann
  • Leopold März
Non-Lectin Papers

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.

Keywords

hyaluronidase Apis mellifera bee venom N-linked carbohydrate chains α1,3-fucosylation 

Abbreviations

GalNAc

N-acetylgalactosamine

GlcNAc

N-acetylglucosamine

PA

pyridylamino

PLA

phospholipase A2

2D-HPLC

two-dimensional HPLC

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Copyright information

© Chapman & Hall 1995

Authors and Affiliations

  • Viktoria Kubelka
    • 1
  • Friedrich Altmann
    • 1
  • Leopold März
    • 1
  1. 1.Institut für Chemie der Universität für Bodenkultur WienViennaAustria

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