Structure of O‐glycosidically linked oligosaccharides synthesized by the insect cell line Sf9

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Abstract
The O‐glycosidically linked Oligosaccharides on the pseudorabies virus (PRV) glycoprotein gp50 synthesized by three different cell lines were studied. The intact membrane protein (gp50) was expressed in Vero cells and in the insect cell line Sf9. In addition, a truncated, secreted form lacking the transmembrane and cytoplasmic domains (gp50T), was expressed in CHO and Sf9 cells. The protein, both in intact and truncated form, synthesized by the two mammalian cells contained only the disaccharide Galβ1‐3GalNAc, either unsubstituted or substituted with one or tow sialic acid residues. By contrast, the major O‐linked structure on gp50 and gp50T synthesized by Sf9 cells was the monosaccharide GalNAc. The Sf9 cells also linked lower amounts of Galβ1‐3GalNAc to gp50 (12%) and gp50T (26%). None of the structures synthesized by Sf9 cells contained sialic acid. Measurements of the two relevant glycosyltransferases revealed that while all three cell lines contain comparable levels of the UDP‐GalNAc:polypeptide, N‐acetylgalactosaminyltransferase activity, there is a greater variation in the levels of UDP‐Gal: N‐acetylgalactosamine, β1‐3 galactosyltransferase, with the Sf9 cells containing the lowest level.