Open AccessPrimary structures of the N‐linked carbohydrate chains from honeybee venom phospholipase A 2
Author(s) -
KUBELKA Viktoria,
ALTMANN Friedrich,
STAUDACHER Erika,
TRETTER Verena,
MÄRZ Leopold,
HÅRD Karl,
KAMERLING Johannis P.,
VLIEGENTHART Johannes F. G.
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb17870.x
Subject(s) - exoglycosidase , chromatography , glycan , chemistry , high performance liquid chromatography , fucose , oligosaccharide , biochemistry , phospholipase , carbohydrate conformation , fractionation , carbohydrate , phospholipase a , mannose , elution , peptide , phospholipase a2 , glycoprotein , polysaccharide , enzyme
The N‐linked carbohydrate chains of phospholipase A 2 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 1 H‐NMR spectroscopy. Ten oligosaccharides consisted of mannose, N ‐acetylglucosamine and fucose α1–6 and/or α1–3 linked to the innermost N ‐acetylglucosamine. Four compounds, which comprised 10% of the oligosaccharide pool from phospholipase A 2 , contained a rarely found terminal element with N ‐acetylgalactosamine. The structures of the 14 N‐glycans from honeybee phospholipase A 2 can be arranged into the following three series: