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In vitro enzymatic activities of ycLeuRS. Deacylation of [3H]-Ile-tRNALeu and [3H]-Met-tRNALeu by 1 μM ycLeuRS (A). Misaminoacylation of yctRNALeu with 25 μM [3H]-methionine (422 μCi/mL) by 1 μM LeuRS (B). ATP hydrolysis in absence (C) or presence (D) of tRNA. The reactions contained 250 μM ATP (600 μCi/mL), amino acids (2.5 mM leucine or 100 mM isoleucine or methionine) and 1 μM ycLeuRS (C). Assays are described in detail in the Supporting Information.
