Assay of galactosyltransferase by high-performance liquid chromatography.
Galactosyltransferase catalyzes transfer of galactose from UDP-galactose to glucose or N-acetylglucosamine with resultant formation of galactosides and UDP. In this new assay galactosyltransferase activity is measured by determining UDP by isocratic high-performance liquid chromatography on an amino-bonded column monitored spectrophotometrically. Concurrently, unreacted UDP-galactose and breakdown products arising from UDP-galactose (UMP and uridine) are also determined. The new technique does not require radioactive substrates, permits usage of saturating concentrations of UDP-galactose, and provides monitoring of side reactions.
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005700:Galactosyltransferases / Q000032:analysis* / Q000097:blood; D006801:Humans; D008892:Milk / Q000201:enzymology; D013056:Spectrophotometry, Ultraviolet; D014529:Uridine / Q000032:analysis; D014530:Uridine Diphosphate / Q000032:analysis; D014531:Uridine Diphosphate Galactose / Q000378:metabolism; D014542:Uridine Monophosphate / Q000032:analysis
D014531:Uridine Diphosphate Galactose; D014530:Uridine Diphosphate; D014542:Uridine Monophosphate; D005700:Galactosyltransferases; D014529:Uridine