Structural characterization of the N-linked oligosaccharides derived from HIVgp120 expressed in lepidopteran cells.

The oligosaccharides of recombinant HIV gp120 expressed in lepidopteran Sf9 cells were analysed after hydrazine release by gel permeation and high pH anion exchange chromatography. N-Linked glycans were exclusively of the oligomannose series and no evidence for charged complex or hybrid type glycans was found. However a glycosylation reaction similar to those found in vertebrates was evident. The major glycoform of gp120, that comprised 30% of all the species analysed, was structurally identified by exoglycosidase digestion and found to be a core fucosylated structure, Manalpha1,6(Manalpha1,3)Manbeta1,4GlcNAc(Fucalpha1+ ++,6)GlcNAc. Further confirmation of the ability of lepidopteran cells to fucosylate N-linked glycans was provided by an in vitro analysis of this reaction using authentic oligosaccharide substrates.