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A capillary zone electrophoresis assay for the nucleoside transfer enzyme adenosine diphosphate-glucose
M W Roberts1, J Preiss, T W Okita
1Institute of Biological Chemistry, Washington State University, Pullman 99163-6340, USA.
Abstract:
Conditions are described for a capillary zone electrophoresis-based assay of ADP-glucose pyrophosphorylase, the first enzyme in the de novo synthesis for alpha-glucan synthesis in plants and bacteria. ATP, ADP-glucose, and other adenine derivatives are effectively resolved and quantified in 12 min of electrophoresis. The method requires minimal sample manipulations and no postincubation hydrolyses, is effective over a wide range of enzyme and substrate concentrations, has a sensitivity comparable to radioisotope assays, and yields reproducible results in either the forward and reverse directions. Moreover, the assay is applicable to other enzymes that utilize nucleosides or their bases as substrates or products.