Enzymic preparation of adenosine 5'-[beta-2P]triphosphate.

A simple method for the preparation of adenosine 5'-[beta-32 P]triphosphate is described. When [32P]orthophosphate was incubated with polyadenylate and phosphoenolpyruvate in the presence of polynucletide phosphorylase (EC 2.7.7.8) and pyruvate kinase (EC 2.7.1.40), up to 75% of 32P radioactivity was recovered in ATP. [32P]ATP was purified to 99.5% radiochemical purity by chromatography on polyethyleneimine-cellulose thin-layer plates. Analysis of hydrolysis products of [32P]ATP with apyrase (EC 3.6.1.5) indicates that 32P in the beta-phosphate position accounts for all 32P label in ATP.