Figure 5.

pH dependence of the phosphate removal from ppp-RNA catalyzed by YvcI. A, YvcI is most active under slight acidic to neutral pH. ppp-RNA (AGU 5S rRNA, 5′-labeled, 0.5 μm) was incubated with YvcI (2 μm) in 1× manganese buffer (2 mm Mn²⁺). The dotted line indicates where different parts of different plates were combined to facilitate interpretation. PEI-cellulose TLC, phosphorimaging of radioactivity is shown. B and C, quantification of orthophosphate (p_i) (B) and pyrophosphate (pp_i) (C) from A, showing means ± S.D. of two independent experiments (technical replicates).