Human blood group glycosyltransferases. I. Purification of n-acetylgalactosaminyltransferase.
An N-acetylgalactosaminyltransferase, which converts blood group O red blood cells to A cells, was purified to homogeneity from plasma of blood group A1 subjects. The enzyme was adsorbed on Sepharose 4B, and after washing out the impurities, the enzyme was eluted with UDP. This procedure resulted in a 70,000- to 100,000-fold increase in specific activity with recovery of about 80%. Further purification of the enzyme was achieved by Bio-Gel P treatment. The final enzyme preparation showed a single protein band, which coincided with enzyme activity, on acrylamide gel electrophoresis, and revealed a single protein band on sodium dodecyl sulfate-gel electrophoresis. Judging from the molecular weight (90,000 to 100,000), which was estimated by Sephadex gel filtration, and the subunit size estimated by sodium dodecyl sulfate-gel electrophoresis, the enzyme is presumably in a dimeric form. The enzyme required Mn2+ and had optimum activity at pH 6.5 to 7.0.
D000017:ABO Blood-Group System; D000116:Acetylgalactosamine; D005700:Galactosyltransferases / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight
D000017:ABO Blood-Group System; D005700:Galactosyltransferases; D000116:Acetylgalactosamine