Porcine A blood group-specific N-acetylgalactosaminyltransferase. I. Purification from porcine submaxillary glands.
The membrane-bound N-acetylgalactosaminyltransferase from porcine submaxillary glands which provides A blood group specificity to mucin has been purified 38,000-fold by affinity chromatography on UDP-hesanolamine-agarose in aqueous Triton X-100. Design of a suitable purification procedure was developed by assessing the strength of interaction between enzyme and affinity adsorbent using batch desorption. The pure transferase has an apparent molecular weight of 100,000 as judged by zonal centrifugation and by sodium dodecyl sulfate polyacrylamide gel electrophoresis in the absence of a reducing agent. The reduced and carboxymethylated protein has an apparent molecular weight of 46,000 and 57,000 as judged by sedimentation equilibrium and sodium dodecyl sulfate polyacrylamide gel electrophoresis, respectively, suggesting that the native enzyme contains two subunits. It is a glycoprotein with a specific activity of 30 micronmol/min/mg of enzyme, which is 55,000 times that reported for the same enzyme isolated from human serum.
D000017:ABO Blood-Group System; D000818:Animals; D002846:Chromatography, Affinity; D006602:Hexosyltransferases / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D013045:Species Specificity; D013363:Submandibular Gland / Q000201:enzymology*; D013552:Swine; D013696:Temperature; D014536:Uridine Diphosphate N-Acetylgalactosamine
D000017:ABO Blood-Group System; D014536:Uridine Diphosphate N-Acetylgalactosamine; D006602:Hexosyltransferases