Fig. 6.
M. synoviae LeuRS CP1 addition to M. mobile LeuRS suppresses mischarging via enhanced pretransfer editing activity. (A) Aminoacylation and misaminoacylation activity of Mm/Ms LeuRS was carried out with 4 μM tRNALeu in the presence of 21 μM [3H]leucine (318 μCi/mL) with 1 μM enzyme or 21 μM [3H]isoleucine (97 μCi/mL) with 1 μM enzyme. Symbols used are as follows: □, Leu-tRNALeu and △, Ile-tRNALeu. (B) The AMP formation activity was measured for Mm/Ms LeuRS using 1 μM enzyme, 10 μM tRNALeu, or tRNALeu with A76 replaced by dideoxyadenosine (ddA-tRNA), 18.1 μM [α-32P]ATP, and 2.5 mM isoleucine. Symbols used are as follows: ◆, reaction without amino acid present (no AA); ▲, isoleucine (Ile); ▼, isoleucine with tRNA (Ile+tRNA); and ■, isoleucine with ddA-tRNA (Ile+ddA). Error bars represent the SD values based on three separate experiments.