title: Purification and properties of N-acetylglucosaminide beta 1----4 galactosyltransferase from embryonal carcinoma cells.
abstract: N-Acetylglucosaminide beta 1----4 galactosyltransferase was chromatographically purified about 1,700-fold from F9 embryonal carcinoma cells after solubilization with Triton X-100, using N-acetylglucosamine as the acceptor. As the last step of the purification, affinity chromatography was performed either on N-acetylglucosamine-Sepharose or on alpha-lactalbumin-Sepharose: in both cases, two protein bands with molecular weights of around 68,000 and 59,000 were detected by SDS-polyacrylamide gel electrophoresis of the purified preparations. The enzymological properties including behavior toward alpha-lactalbumin were very similar to those of the enzyme from other sources. The specificity of the enzyme was confirmed by determining the structure of the product; it was mostly Gal beta 1----4GlcNAc. beta-Galactosidase-treated embryoglycan (poly-N-acetyllactosamine) and asialo-agalactofetuin could serve as acceptors with the purified enzyme. Thus, the embryonic enzyme, apparently involved in the synthesis of poly-N-acetyllactosamines, has properties similar in several respects to those of the beta-galactosyltransferases so far studied.
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D054278:Embryonal Carcinoma Stem Cells; D005700:Galactosyltransferases / Q000032:analysis / Q000302:isolation & purification; D007788:Lactose Synthase / Q000032:analysis / Q000302:isolation & purification*; D008715:Methionine / Q000032:analysis; D051379:Mice; D014411:Neoplastic Stem Cells / Q000201:enzymology*; D013379:Substrate Specificity; D013462:Sulfur Radioisotopes; D014407:Tumor Cells, Cultured
chemical_list: D013462:Sulfur Radioisotopes; D008715:Methionine; D005700:Galactosyltransferases; D007788:Lactose Synthase