AN IMPROVED METHOD FOR THE SYNTHESIS OF GDP-HEXANOLAMINE DERIVATIVES,KEY REAGENTS FOR THE PURIFICATION AND CHARACTERIZATION OF CARBOHYDRATE PROCESSING ENZYMES

As part of our ongoing studies on the isolation and characterisation o f alpha-(1,3)-fucosyltransferase (FucT) enzymes, an improved method fo r the synthesis and purification of the key biological tool GDP-hexano lamine (1a), together with detailed experimental conditions and produc t characterisation are reported. We demonstrate the viability of the p roduct obtained, by its competitive inhibition of an alpha-(1,3)-fucos yltransferase. Subsequently, we are able to use it to derivatise a sep harose gel column which we show is highly efficient in the purificatio n of a recombinant alpha-(1,3)-fucosyltransferase from a crude prepara tion. The GDP-hexanolamine is converted to the photoaffinity probe GDP -hexanolaminyl-4-azidosalicylate (1b), useful in the characterisation of fucosyltransferase enzymes.