Video 2. Förster resonance energy transfer (FRET) assay on supported lipid bilayers (SLBs).

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PBP1B^Ec was reconstituted in E. coli polar lipids SLBs at a 1:10⁵ (mol:mol) protein-to-lipid ratio along lipid II-Atto647 and lipid II-Atto550. Membranes were incubated with 5 µM lipid II in the presence or absence of 1 mM ampicillin. To detect FRET, the fluorescence of the acceptor Atto647n was bleached within a region. In the subsequent frame, the fluorescence of Atto550 increased, indicating the presence of FRET. In the presence of ampicillin, this increase did not happen.