HPLC METHOD FOR THE DETERMINATION OF FUC TO ASN-LINKED GLCNAC FUCOSYL-TRANSFERASES

Mammalian cells often contain an enzyme which transfers fucose onto th e reducing terminal GlcNAc (GlcNAc-1) of N-glycans with an alpha 1,6-l inkage. In plants, on the other hand, the fucose is transferred to Glc NAc-1 with an alpha 1,3-linkage. Insect cells can exhibit both enzymat ic activities. Hitherto, the activity of these fucosyltransferases has been determined by the incorporation of radioactively labelled fucose into an acceptor glycopeptide. This assay, however, cannot discrimina te these two activities. Here we report on the use of dansylated glyco asparagine for the specific determination of 1,3- and 1,6-fucosyltrans ferases. The two possible products and the substrate are separated on a reversed phase column and detected by fluorescence.