N-acetylgalactosaminyl disaccharide terminals contained in presumed carbohydrate epitopes specific to sea-squirt allergy

Using the methods of methylation/GLC, MS, and 1H-NMR analyses, a disaccharide structure of GalNAc alpha 1----2Fuc1---- was identified as one of the major structural units at non-reducing terminals of a highly branched and N-glycosidically linked carbohydrate on an allergenically active glycopeptide, Gp-2 (Mr ca. 8,000). Gp-2 was one of the major fractions in the Pronase digest of a sea-squirt antigen, Gi-rep, and is capable of eliciting the skin reaction specific to patients with sea-squirt allergy similarly to Gi-rep. An additional disaccharide structure of GalNAc1----(3/4)HexNAc1---- was also expected as the other major non-reducing terminal unit, though the anomeric configuration of the terminal GalNAc could not be identified. As Gp-2 carbohydrate was found to contain 3 mol each of the two types of terminal GalNAc-containing units, both were nominated as possible components of the IO4- oxidation-sensitive epitopes which are responsible for the allergenic activity in Gp-2 and its mother antigen, Gi-rep. A few moles of Fuc and Gal were also detected as minor non-reducing terminals in addition to the 6 mol of the GalNAc-containing units.