Use of Crithidia fasciculata extract for the facile enzymatic synthesis of GDP-L-3HFucose

For studies involving glycosyltransferases and nucleotide sugar transporters, radioactive nucleotide sugars are critical reagents. Of these, GDP-L-[3H]Fucose is currently commercially unavailable. Here, we present a facile approach for the preparation of GDP-[3H]-L-Fucose, using the enzymatic machinery present in the cytosol of the non-infectious and easily cultivated protozoan, Crithidia fasciculata, and its purification by solid phase extraction ion exchange chromatography.For studies involving glycosyltransferases and nucleotide sugar transporters, radioactive nucleotide sugars are critical reagents. Of these, GDP-L-[3H]Fucose is currently commercially unavailable. Here, we present a facile approach for the preparation of GDP-[3H]-L-Fucose, using the enzymatic machinery present in the cytosol of the non-infectious and easily cultivated protozoan, Crithidia fasciculata, and its purification by solid phase extraction ion exchange chromatography.