Analysis of glucose 1-phosphate at the picomole level with [C]UTP and uridine 5′-diphosphoglucose pyrophosphorylase |
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Authors: | Chi P CheungRobert J Suhadolnik |
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Institution: | Department of Biochemistry, Temple University School of Medicine, Philadelphia, Pennsylvania 19140 USA |
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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. |
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