Specificity of submaxillary gland sialyltransferases.
A method has been developed to determine the activities of specific sialyltransferases by analysis of the products of the reaction. This method, which utilizes high performance liquid chromatography, distinguishes addition of sialic acid to the N-acetylgalactosamine vs. galactose residues of the mucin disaccharide Gal beta(1 leads to 3)GalNac, and can be used to distinguish formation of the 3'- and 6'-isomers of sialyllactose. For the bovine, ovine, and porcine submaxillary extracts, more than 95% of the activity with asialo ovine submaxillary mucin is due to formation of NeuAc alpha(2 leads to 6)GalNAc. With lactose as the acceptor, more than 95% of the alpha(2 leads to 3) isomer is produced. Activity with asialofetuin is due solely to the O-linked chain, with relative activity toward the galactose vs. GalNAc residues of 0.32, 1.5, and 0.10 for bovine, ovine, and porcine, respectively. The rat submaxillary gland extract showed equal formation of 3'- and 6'-sialyllactose, and very low activity with asialo ovine submaxillary mucin. However, at least 40% of the activity toward the Gal beta(1 leads to 3)GalNAc disaccharide of asialofetuin was directed toward the GalNAc residue. The relative preference of the N-acetylgalactosaminide alpha(2 leads to 6) sialyltransferase for a monosaccharide vs. a substituted GalNAc may play a role in regulation of chain length during mucin synthesis.
D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D066298:In Vitro Techniques; D051381:Rats; D012756:Sheep; D012799:Sialyltransferases / Q000378:metabolism*; D013045:Species Specificity; D013363:Submandibular Gland / Q000201:enzymology*; D013379:Substrate Specificity; D013552:Swine; D014166:Transferases / Q000378:metabolism*
D014166:Transferases; D012799:Sialyltransferases