FIG. 5.
Kinetics of ATP hydrolysis. (A) Reaction mixtures containing 50 mM Tris-HCl (pH 7.5), 5 mM DTT, 1 mM MnCl2, 0.2 mM [γ-32P]ATP, and cvRTP as specified (in nanograms per 10 μl) were incubated at 37°C. Aliquots (10 μl) were withdrawn at times indicated and quenched immediately with formic acid. (B) Reaction mixtures (110 μl) containing 50 mM Tris-HCl (pH 7.5), 1 mM MnCl2, 5 mM DTT, 0.2 mM [γ-32P]ATP or [α-32P]ATP as specified, and 1.1 μg of cvRTP were incubated at 37°C. Aliquots (10 μl) were withdrawn at times indicated and quenched immediately with formic acid. The reaction products were analyzed by TLC. The extent of 32Pi, [α32P]ADP, or [α32P]AMP formation (from 2 nmol of input ATP per sample) is plotted as a function of time.