Lipid I and II formation with cell-free expressed Mur enzymes and Bs-MraY·nanodisc (DMPG) complexes.A, SDS-PAGE analysis of purified soluble Mur proteins. Samples were separated by 11% Tricine gels and stained with Coomassie Blue. B, TLC detection of lipid I and lipid II formation by cell-free expressed Mur enzymes and Bs-MraY·nanodisc (DMPG) complexes. Similar results were obtained in corresponding combinations with cell-free expressed Ec-MraY·nanodisc (DMPG) complexes (data not shown). As controls, in vivo expressed Mur enzymes of S. aureus and cell-free expressed Bs-MraY·nanodisc (DMPG) complexes were used for lipid I and lipid II formation.