Fig. 4: Potential TAG entry sites to the central cavity and cavity analysis.

a Side view of lateral clefts TM5-TM8^OUT (left) and TM2-TM11^OUT (right) in outward-facing conformation of MHAS2168 crystal structure. MHAS2168 color scheme same as in Fig. 1b. Point mutation sites are shown as colored sticks. Linker helices A and B are depicted partially transparent. b Side view of lateral clefts TM5-TM8^IN (left) and TM2-TM11^IN (right) in inward-facing conformation of MHAS2168 homology model. MHAS2168 and point mutation sites’ color scheme same as in panel a. c Enlarged periplasmic top view of the central cavity and lateral clefts in outward-facing conformation of MHAS2168 crystal structure. MHAS2168 and point mutation sites’ color scheme same as in panel a. d Hydrophobicity surface of the C-domain central cavity wall of MHAS2168 outward-facing crystal structure with mutated residues shown as colored sticks. Hydrophobicity color scheme: hydrophobic – gold; hydrophilic – cyan. e Vancomycin sensitivity assays in M. smegmatis dKO cells, complemented with wild type LprG/Rv1410 operon (WT_Mtb), or mutant operons containing unaltered LprG (Rv1411) and mutated transporter Rv1410 as indicated. f Analogous analysis as in (e), with M. smegmatis dKO cells expressing instead wild type MHAS2167/68 operon (WT_MH) or mutant operons containing unaltered LprG (MHAS2167) and mutated transporter MHAS2168 as indicated. The growth curves in (e) and (f) are representative of three biological replicates and data are presented as mean values +/- SD of four technical replicates.