The influence of quaternary structure on glycosylation was evaluated in a macrophage-like cell line, P388D1. This cell line simultaneously synthesizes two structurally related glycoproteins, Mac-1 and LFA-1. Mac-1 and LFA-1 each contain two subunits in non-covalent association in an α1β1 structure. The β-chain polypeptides of these two glycoproteins have identical primary structures while their α-chain polypeptides are distinct. For both Mac-1 and LFA-1, the association of the α- and β-chains occure prior to any Golgi-mediated processing of the oligosaccharide moieties on either one of the subunits. To evaluate the effects of differential subunit association on the site-specific glycosylation of the β-chain, [3H]glucosamine-labeled oligosaccharides were isolated from the β-chain of Mac-1 and LFA-1 and were compared by a variety of enzymatic and chromatographic techniques. Reverse-phase high performance liquid chromatography analyses of tryptic-chymotryptic glycopeptides suggest that each β-chain has at least five glycosylation sites. Structural analysis of oligosaccharides from each corresponding glycopeptide fraction of the β-chains of Mac-1 or LFA-1 (comparing their glycosidase sensitivities, behavior on serial lectin affinity chromatography, size heterogeneity, extent of sialylation, and branching) indicates that the LFA-1 β-chain is glycosylated substantially differently on at least four of its sites, compared to the corresponding sites of the Mac-1 β-chain, even though they are simultaneously synthesized in the same cells. Thus, these data demonstrate that quaternary structure can influence the site-specific glycosylation of a protein, even when the polypeptide structure and the cellular glycosylation machinery remains constant.
|Original language||English (US)|
|Number of pages||11|
|Journal||Journal of Biological Chemistry|
|State||Published - 1986|
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