Occurrence of alpha-D-galactosyl residues in the thyroglobulins from several species. Localization in the saccharide chains of the complex carbohydrate units.
Treatment of thyroglobulins from several mammalian sources (calf, sheep, pig, dog, rat, rabbit, guinea pig, and man) with alpha-galactosidase demonstrated a species-dependent occurrence of terminal alpha-D-galactosyl residues which ranged from 11 mol/mol of protein (23% of total galactose) in calf to a complete absence in man. The presence of the alpha-D-galactosyl groups resulted in a partial binding of the thyroglobulins (greater than 70% in calf and sheep) to Bandeiraea simplicifolia I-agarose, and this lectin-thyroglobulin interaction could be quantitated by a solid-phase assay utilizing 125I-labeled B. simplicifolia I. Sequential glycosidase digestions of calf thyroglobulin glycopeptides containing the complex carbohydrate unit (unit B) and characterization of oligosaccharide obtained by partial acid hydrolysis indicated that the alpha-D-galactosyl residues are located on oligosaccharide branches with an alpha-D-Gal-(1----3)-beta-D-Gal-(1----4)-D-GlcNAc sequence. While mild acid treatment of calf thyroglobulin glycopeptides yielded a disaccharide, alpha-D-Gal-(1----3)-D-Gal, and a trisaccharide, alpha-D-Gal-(1----3)-beta-D-Gal-(1----4)-D-GlcNAc, which could be resolved by B. simplicifolia I-agarose or thin-layer chromatography, similar hydrolysis of the human unit B-containing glycopeptides did not produce such components. A study of various glycopeptides indicated that the alpha-D-galactosyl residues are unevenly distributed among the multiple complex carbohydrate units of calf thyroglobulin and are preferentially located in units with a relatively low sialic acid content. During affinity chromatography on B. simplicifolia I-agarose, glycopeptides with multiple alpha-D-galactosyl groups bound more firmly to the lectin than those which contained only a single residue. In contrast to the alpha-D-galactosyl residues, beta-linked galactose of calf thyroglobulin was primarily bound in penultimate locations being susceptible to enzymatic release only after prior removal of capping sialyl and alpha-D-galactosyl groups. The isolation of N-acetyllactosamine and a beta-D-Gal----beta-D-GlcNAc----D-Man trisaccharide from partial acid hydrolysates helped to position the beta-D-galactosyl residues in the oligosaccharide branches of the complex carbohydrate units.
D000818:Animals; D002236:Carbohydrate Conformation; D002417:Cattle; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002855:Chromatography, Thin Layer; D004285:Dogs; D005690:Galactose / Q000032:analysis*; D006168:Guinea Pigs; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008322:Mammals / Q000378:metabolism*; D037121:Plant Lectins; D011817:Rabbits; D051381:Rats; D012756:Sheep; D013552:Swine; D013954:Thyroglobulin / Q000032:analysis*; D013961:Thyroid Gland / Q000032:analysis; D000519:alpha-Galactosidase / Q000378:metabolism
C045475:Griffonia simplicifolia lectins; D037102:Lectins; D037121:Plant Lectins; D013954:Thyroglobulin; D000519:alpha-Galactosidase; D005690:Galactose