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. 2017 Jul 29;45(16):9573–9582. doi: 10.1093/nar/gkx673

Figure 3.

Figure 3.

In vitro binding of MTM and PKM to the NPET. (A and B) DMS probing of interactions of antibiotics with (A) Escherichia coli or (B) Staphylococcus aureus ribosomes. Cladinose-containing macrolide ERY, ketolide TEL or PTC-binding antibiotic CHL were used as controls. The arrows indicate the 23S rRNA residues A2058 and A2059, located in the macrolide-binding site in the NPET. Samples in the lanes marked ‘none’ contained no antibiotic. (C) Competitive binding of non-radioactive ERY, PKM or MTM with [14C] ERY to E. coli 70S ribosomes. The MTM and PKM binding experiments were done in triplicates, the binding of the control antibiotic, ERY, was analyzed in duplicate samples. Assuming ERY Kd being 10.8 nM (18), the estimated Ki of MTM and PKM were 13.1 ± 3.5 and 3.2 ± 1.6 μM, respectively.