The major oligosaccharides in the large subunit of the hemagglutinin from fowl plague virus, strain Dutch. Structure elucidation by one-dimensional and two-dimensional 1H nuclear magnetic resonance and by methylation analysis

Abstract

The N‐glycosidically linked glycans in the large subunit (HA₁) of the hemagglutinin from fowl plague virus, strain Dutch (containing about 15%, w/w, of carbohydrates), were liberated by alkaline hydrolysis, and were filtrated through Bio‐Gel as the re‐N‐acetylated oligosaccharide alditols. One major fraction (90%, mol/mol) was obtained. It was subfractionated by concanavalin A affinity chromatography and was analyzed by methylation/capillary gas chromatography/mass fragmentography and especially by one‐dimensional and two‐dimensional ¹H nuclear magnetic resonance. The major HA₁ glycans, which are not sialylated, were thus found to comprise about 40%, 30% and 20% (mol/mol), respectively, of biantennary intersected, biantennary, and triantennary N‐acetyllactosaminic (‘complex’) oligosaccharides. About two thirds of the internal GlcNAc residues in these glycans are substituted by Fuc(α1→6), all the triantennary species carry the third Gal(β1→4)GlcNAc(β→ unit at the Man(α→6)‐branch, and roughly one fourth of the N‐acetyllactosamine units in the non‐intersected biantennary oligosaccharides are incomplete.