Primary structures of the N-linked carbohydrate chains from honeybee venom phospholipase A2.
The N-linked carbohydrate chains of phospholipase A2 from honeybee (Apis mellifera) were released from glycopeptides with peptide-N-glycanase A and reductively aminated with 2-aminopyridine. The fluorescent derivatives were separated by size-fractionation and reverse-phase HPLC, yielding 14 fractions. Structural analysis was accomplished by compositional and methylation analyses, by comparison of the HPLC elution patterns with reference oligosaccharides, by stepwise exoglycosidase digestions which were monitored by HPLC, and, where necessary, by 500-MHz 1H-NMR spectroscopy. Ten oligosaccharides consisted of mannose, N-acetylglucosamine and fucose alpha 1-6 and/or alpha 1-3 linked to the innermost N-acetylglucosamine. Four compounds, which comprised 10% of the oligosaccharide pool from phospholipase A2, contained a rarely found terminal element with N-acetylgalactosamine. The structures of the 14 N-glycans from honeybee phospholipase A2 can be arranged into the following three series: [formula: see text]
Institut für Chemie, Universität für Bodenkultur, Vienna, Austria.
V
V
Kubelka
F
F
Altmann
E
E
Staudacher
V
V
Tretter
L
L
März
K
K
Hård
J P
JP
Kamerling
J F
JF
Vliegenthart
D000818:Animals; D001514:Bee Venoms / Q000737:chemistry*; D001516:Bees; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D010741:Phospholipases A / Q000737:chemistry*; D054467:Phospholipases A2
D001514:Bee Venoms; D009844:Oligosaccharides; D010741:Phospholipases A; D054467:Phospholipases A2