AN APPROACH FOR FLUOROMETRIC-DETERMINATION OF GLYCOSYLTRANSFERASE ACTIVITIES

A new strategy for the fluorometric determination of glycosyltransfera se activities is reported. The method involves dansyl chloride derivat ization of the reduced form (pNH(2)phenyl) of a hydrophobic, aglycon m oiety covalently linked to a number of acceptor substrates (pNO(2)phen yl). Focusing on the Golgi enzyme core 2 N-acetylglucosaminyltransfera se, we found that synthesis and fractionation of the dansylated substr ate derivative were rapid, easy and inexpensive. Additionally, the cor responding enzyme assay proved reproducible and very sensitive, as 0.4 pmol of reaction product were readily detected. This fluorometric app roach appears therefore to be a valid tool for investigating the monit oring differential expression of glycosyltransferases exhibiting low l evels of enzyme activity.