FIGURE 4.
Characterization of the RluD-directed pseudouridine synthesis. Determination of catalytic parameters of RluD for substrate 50S subunits. The initial rate of pseudouridylation was measured in the presence of 190 nM purified RluD and increasing concentrations of ΔrlmH/ΔrluD 50S subunits. RlmH-dependent pseudouridine methylation was used as the reporter reaction for monitoring pseudouridine synthase activity of RluD in a coupled enzyme assay. The amount of [3H]-methyl groups incorporated was monitored by acid precipitation and scintillation counting. One experiment representative of three repeats is shown.
