Fig. 4.
Mutations in aspS are associated with TBA161-C resistance. (A) Susceptibility of M. marinum wild type (Mmar) and TBA161-C-resistant isolates (Mmar-R-TBA161-C) towards TBA161-C after 4 days of incubation. (B) Susceptibility of Mtb wild type and TBA 161-resistant isolates (Mtb-R-TBA161-C) towards TBA161-C was measured after 7 days. (C) M. marinum wild type transformed with pMS2-aspSMtb (Mmar+aspSMtb), and pMS2-aspSMtbF526L (Mmar+aspSMtbF526L) were incubated with compound TBA161-C for 4 days at the indicated concentrations. (D) Mtb carrying plasmids pMS2-aspSMmar (Mtb+aspSMmar) and pMS2-aspSMmarR168G (Mtb+aspSMmarR168G) were incubated with twofold dilutions of compound TBA161-C for 7 days. (E) TBA161-C (orange) docked into the catalytic subdomain of chain A of Mtb AspS (PDB ID: 5W25). The zoom-in shows TBA161-C in stick representation, together with AspS residues aligning the binding pocket. These include R171 (blue), the three residues of which side chains were treated flexibly during docking (grey), and T570 of chain B (dark brown). The distant F526 residue is shown in blue. For clarity, the L200 label and all hydrogen atoms are omitted. Data are mean of duplicatesĀ±s.d.