A rapid and sensitive assay for quantifying the activity of both aerobic and anaerobic ribonucleotide reductases acting upon any or all substrates
Fig 8
Assay data with class Ia E. coli ribonucleotide reductase measuring the reduction of all four ribonucleotide substrates simultaneously.
(A) Activity of EcNrdA in the presence of substrates, effectors, and varying amounts of dATP. (B) Activity of EcNrdA in the presence of substrates, effectors, and varying amounts of dGTP. (C) Activity of EcNrdA in the presence of substrates, effectors, and varying amounts of TTP. Each bar is representative of the average activity +/- standard error of the mean for three replicates. The values of each bar are shown in Table 2. Except for the indicated concentrations of allosteric effectors on the X-axis, components and concentrations were as follows: Trx (30 μM), TrxR (0.5 μM), NADPH (200 μM), CDP (70 μM), UDP (50 μM), GDP (200 μM), ADP (110 μM), NrdA (0.1 μM dimer), NrdB (0.5 μM dimer).