Identification of a alpha-NeuAc-(2----3)-beta-D-galactopyranosyl N-acetyl-beta-D-galactosaminyltransferase in human kidney.
Microsomal preparations from human kidney were found to contain enzymic activity capable to transfer N-acetylgalactosamine from UDP-N-acetylgalactosamine to native bovine fetuin. The acceptor structures on the fetuin molecules were identified as N- as well as O-linked glycans with a markedly higher incorporation into the N-linked carbohydrate chains. Analysis of the alkali-labile transferase products by thin-layer chromatography indicated that the enzyme is able to synthesize structures having mobilities identical with those found on glycophorin from Cad erythrocytes. Mild acid treatment and enzymic hydrolysis with N-acetylhexosaminidase from jack beans of the N-linked transferase products suggested that beta-D-GalpNAc-(1----4)-[alpha-NeuAc-(2----3)]-beta-D-Galp-(1----s tructures were formed by the enzymic reaction on both N- and O-linked acceptors. The enzyme might, therefore, be involved in the biosynthesis of Sda (and Cad) antigenic structures. By use of various oligosaccharides, glycopeptides, and glycolipids having well characterized carbohydrate sequences, the acceptor-substrate specificity of the N-acetylgalactosaminyltransferase was determined. The enzyme generally recognized alpha-NeuAc-(2----3)-beta-D-Gal groups as acceptors, but in a certain conformation. Thus, tri- and tetra-saccharide alditols, native human glycophorin A, and GM3 were not acceptor substrates although they carry the potential disaccharide acceptor unit. When these structures were presented as sialyl-(2----3)-lactose or as a tryptic peptide from glycophorin A, they were shown to be rather good acceptor substrates for the N-acetyl-beta-D-galactosaminyltransferase from human kidney.
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002250:Carbon Radioisotopes; D002417:Cattle; D005700:Galactosyltransferases / Q000378:metabolism*; D006017:Glycolipids; D006023:Glycoproteins; D006801:Humans; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D017350:N-Acetylgalactosaminyltransferases; D013379:Substrate Specificity; D000509:alpha-Fetoproteins
D002250:Carbon Radioisotopes; D006017:Glycolipids; D006023:Glycoproteins; D000509:alpha-Fetoproteins; D005700:Galactosyltransferases; D017350:N-Acetylgalactosaminyltransferases; C049758:N-acetylneuraminyl(2-3)galactopyranosyl N-acetylgalactosaminyltransferase