Figure 3. Amphiphilic cavity characterization.
A. A model of lipid-linked-disaccharide docked into the amphiphilic cavity of LtaA (see also Extended Data Fig. 7A). The lipid tails length corresponds to C16 chains19. Right inlet, Top view of anchor-LLD binding pocket and residues coordinating its gentiobiosyl headgroup. B. Representative traces of LtaA catalyzed flipping in the presence of different disaccharides at given concentrations (n ≥ 3). C. Relative flipping activity of assays showed in B. Error bars indicate the s.d. of technical replicates, n ≥ 3. D. Cells growth on IPTG-free C+Y agar plates at 37°C and in the presence of 5% CO2. ΔltaA represents the S. aureus NCTC8325 ΔltaA mutant; LtaA WT represents ΔltaA mutant complemented with wild type ltaA on a multicopy vector (pLOW); Ctrl vector indicates ΔltaA mutant complemented with pLOW carrying a nonrelated gene (dcas9); the other labels represent ΔltaA mutant complemented with ltaA with corresponding point mutations. E. Representative traces of flipping activity of LtaA-WT and variants. F. Relative flipping activity of assays showed in E. Asterisk marks addition of dithionite. Error bars indicate the s.d. of technical replicates, n ≥ 3. Source data for B-C and E-F are available with the paper online.