Direct demonstration of heterogeneous, sulfated O-linked carbohydrate chains on an endothelial ligand for L-selectin.

We have previously identified endothelial ligands for L-selectin as sialylated, fucosylated and sulfated glycoproteins of approximately 50 kDa and 90 kDa (Sgp50 and Sgp90). In this report, we use the beta elimination reaction to demonstrate directly the presence of sulfated O-linked sugar chains on one of these ligands, after metabolic labeling with radiolabeled sulfate or fucose. All of the sulfated and the majority of the fucosylated O-linked sugar chains were shown to be sialylated by affinity chromatography on a Limax agglutinin column. Analyses by anion exchange and gel permeation chromatography revealed a complexity of sugar chains, which were heterogeneous both in charge and size. Charged groups other than sialic acid appeared to exert a predominant influence on the total charge of the sugar chains. The probable existence of a varying number of sulfate modifications per sugar chain is discussed.