Fig. 1: Activity and general structural features of S. aureus TarGH.

a 2D chemical structure of the lipid-linked WTA precursor substrate of S. aureus TarGH. b Cartoon schematic of WTA precursor transport, peptidoglycan attachment and C₅₅-P recycling within S. aureus. c ATPase activity of S. aureus TarGH expressed in E. coli or L. lactis measured using a malachite green assay. Data points represent the mean of three independent samples ± the standard deviation while Vmax error is reported within the 95% confidence interval. d Cryo-EM reconstruction of E. coli expressed S. aureus TarGH (2.3 Å resolution) bound to ATPγS viewed as heterotetrametric (top) or heterodimeric half-transporter (bottom) assemblies. Membrane spanning TarG and nucleotide binding domain TarH dimers are coloured light/dark blue and green (top, left), by electrostatic potential surface depiction (middle), sequence conservation (Consurf) (bottom, left), or a cartoon depiction with one monomer of each coloured rainbow from N- (blue) to C- (red) terminus (right). Bound LMNG and ATPγS molecules are shown as spheres with heteroatom colouring (carbons purple or grey, respectively) and transmembrane (TM) helices, reentrant helices (EH), interfacial helices (IF1/2) and extracellular loop (EL) of TarG are labelled along with the gate motif (GM) and gate helix (α1) of TarH. Vertical disposition of the inter-TarG aromatic constriction is denoted by a grey dashed line.