Inhibition of glycosyltransferase activities as the basis for drug development.

Glycosyltransferases are involved in the biosynthesis of protein-bound glycan chains that have multiple and important biological functions in all species. In this protocol, we describe methods to assess the inhibition of glycosyltransferase activities. The kinetic mechanisms of the enzymes, information from structural studies and preliminary inhibition studies can aid in designing appropriate inhibitors. The inhibition of beta4-Gal-transferase can be studied with GlcNAc derivatives that act as alternative acceptor substrate analogs and are expected to dock in the acceptor binding site of the enzyme. The inhibition of core 2 beta6-GlcNAc-transferase can be studied with compounds that may compete with binding of the acceptor or glycosyl-donor substrate. Another example is the use of a class of amino acid specific reagents as inhibitors that help to obtain information about amino acid residues at or near the active site of dolichol-phosphate-mannose synthase or those involved in the enzyme mechanism. These inhibitors can be useful for studies of glycan functions, and have potential as therapeutic drugs for a number of diseases involving glycosylation.