Analysis of glucose 1-phosphate at the picomole level with [ 14C]UTP and uridine 5′-diphosphoglucose pyrophosphorylase

Abstract

A new sensitive method is described for glucose 1-phosphate analysis. The key reaction is the pyrophosphorolysis of UDP-glucose catalyzed by uridine 5′-diphosphoglucose pyrophosphorylase. The reaction product, [ 14C]UDP-glucose, is separated from [ 14C]UTP by adsorbing [ 14C]UTP selectively onto polyethyleneimine cellulose or by separating both labeled compounds on one-dimensional polyethyleneimine thin-layer chromatograms. The sensitivity of the method for glucose 1-phosphate analysis is 5 pmol. The method has been successfully employed to monitor the level of glucose 1-phosphate in early germination of wheat embryos.