D. Sykes, S. S. Rana, J. Barlow
Feb 1, 1983
Citations
0
Influential Citations
7
Citations
Journal
Carbohydrate Research
Abstract
Abstract Starting from phenyl 2-acetamido-2-deoxy-4,6- O -( p -methoxybenzylidene)-β- d -glucopyranoside ( 1 ), chemical syntheses were developed for phenyl 2-acetamido-2-deoxy-3- O -β- d -galactopyranosyl-β- d -glucopyranoside ( 4 ) and phenyl 2-acetamido-2-deoxy-4- O -α- l -fucopyranosyl-3- O -β- d -galactopyranosyl-β- d -glucopyranoside ( 8 ). Thin-layer chromatography in the solvent system 6:4:1:5 (v/v) 2-propanol-ethyl acetate-ammonium hydroxide-water clearly separated the synthetic trisaccharide 8 ( R F 0.69) from synthetic disaccharide 4 ( R F 0.78), fucose ( R F 0.56), and GDP-fucose (which remained at the origin). Based upon this observation, a modified method for the determination of GDP- l -fucose: N -acetylglucosaminide-(1→4)-α- l -fucosyltransferase was developed that employed the synthetic disaccharide 4 as an acceptor, and compound 8 as an authentic reference-compound. This modified assay-procedure can simultaneously monitor possible competing reactions which may interfere with determination of α-(1→4)- l -fucosyltransferase activity; these include phosphorylase and α- l -fucosidase activities, and incorporation of α- l -[ 14 C]-fucose into endogenous acceptors of enzyme preparations. Thus, the modified assay-procedure should facilitate determination of α-(1→4)- l -fucosyltransferase.