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Enzymatic analyses of HisRS and its CMT2W mutants. (A) In vitro tRNA aminoacylation assay with purified HisRS proteins. A reaction with WT HisRS, but no tRNA, was included as a control. (B) Steady-state kinetics of WT and mutant HisRS proteins for histidine and ATP determined by ATP/PPi exchange analysis. Data are presented as mean ± SD (n = 3 replicates per group). kcat, catalytic rate constant; N/D, not determined.
