The asparagine-linked carbohydrate of honeybee venom hyaluronidase.
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 alpha 1,3-monofucosylated or alpha 1,3-(alpha 1,6-)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 GalNAc beta 1-4[Fuc alpha 1-3]GlcNAc beta 1- in beta 1,2-linkage to the core alpha 1,3-mannosyl residue. No evidence for the presence of O-glycans or sialic acids could be found.
Kubelka
V
V
Institut für Chemie, Universität für Bodenkultur Wien, Austria.
Altmann
F
F
März
L
L
D000631:Aminopyridines / Q000737:chemistry; D000818:Animals; D001216:Asparagine / Q000032:analysis / Q000737:chemistry; D001514:Bee Venoms / Q000201:enzymology*; D002240:Carbohydrate Sequence; D002417:Cattle; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange / Q000379:methods; D006821:Hyaluronoglucosaminidase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D008361:Mannosidases / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry; D012794:Sialic Acids / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism
D000631:Aminopyridines; D001514:Bee Venoms; D009844:Oligosaccharides; D011134:Polysaccharides; D012794:Sialic Acids; D001216:Asparagine; D008361:Mannosidases; D006821:Hyaluronoglucosaminidase; D014357:Trypsin