Fig. 5
From: Coordination of capsule assembly and cell wall biosynthesis in Staphylococcus aureus
Attachment of CP to the ultimate PG building block lipid IIPG is mediated by LcpC. a [14C]lipid Icap and lipid IIPG or b lipid Icap and [14C]lipid IIPG were incubated with LcpC in the absence or presence of CapA1. The resulting reaction product (LIIPG-CP; bold) exhibits an altered migration behavior on the TLC compared to the individual substrates, lipid IIPG and lipid Icap. PBP2 catalyzes the transglycosylation of the LcpC reaction product LIIPG-CP (lane 4) resulting in the polymerization of radiolabeled lipid IIPG-CP into hydrophilic glycan strands that retain in the water phase and are not extracted after BuOH/PyrAc treatment resulting in disappearance of the corresponding band on the chromatogram. The PBP2 catalyzed conversion is inhibited in the presence of the antibiotic moenomycin (MOE; 10 µM) (B, lane 5). c Quantification of lipid Icap attachment to lipid IIPG. Radiolabeled bands were quantified by phosphoimaging. Experiments were performed in triplicate. The error bars represent the ± SD from three biological replicates. d LcpC catalyzes the attachment of lipid IIcap (lane 3) and the ultimate CP lipid intermediate lipid IIIcap (lane 4) and the final reaction product is processed by PBP2 added posterior to the LcpC reaction (lane 5). Representative TLC of 2 independent experiments
