Partial characterization of the oligosaccharides of mouse thymocyte Thy-1 glycoprotein.
Four glycopeptides (I, IIA, IIB, III) with different oligosaccharide structures were isolated from purified mouse thymocyte Thy-1 glycoprotein. The glycoprotein was digested with Pronase, and the glycopeptide fraction was isolated by gel filtration and acetylated with [3H]acetic anhydride. The different glycan structures were separated by affinity chromatography on concanavalin A-Sepharose 4B and lentil lectin-Sepharose 4B. Size determinations of intact and exoglycosidase- and endoglycosidase-digested glycopeptides were performed by gel filtration on Bio-Gel P-6, calibrated with glycopeptides of known structure. On the basis of these experiments and on the behaviour of the glycopeptides on the lectin columns, the following structures of the oligosaccharide chains were proposed: I, triantennary 'complex-type' with terminal fucose; IIA, biantennary 'complex-type' without fucose; IIB, biantennary 'complex-type' with fucose; III, a mixture of 'high-mannose' chains containing either five or six mannose residues (approx. 50% of each). Amino acid analysis of the glycopeptides showed that the predominant oligosaccharide at glycosylation-site Asn-23 was of 'high-mannose' type, whereas the other two sites (Asn-75 and Asn-99) were glycosylated with 'complex-type' chains. Both these sites were shown to be variably glycosylated. The major glycans linked to Asn-75 were of structures I and IIB, whereas all three 'complex-type' chains were represented at Asn-99. The results presented explain the previously reported carbohydrate heterogeneity of thymocyte Thy-1 glycoprotein.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000954:Antigens, Surface; D002236:Carbohydrate Conformation; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D006020:Glycopeptides / Q000032:analysis; D006026:Glycoside Hydrolases; D051379:Mice; D009844:Oligosaccharides / Q000032:analysis*; D018800:Thy-1 Antigens; D013950:Thymus Gland / Q000032:analysis* / Q000166:cytology
D000596:Amino Acids; D000954:Antigens, Surface; D006020:Glycopeptides; D009844:Oligosaccharides; D018800:Thy-1 Antigens; D006026:Glycoside Hydrolases