Fig. 2: Assay for BAM function in native environment.
From: High-throughput screening of BAM inhibitors in native membrane environment
a Cryo-electron micrograph of 50 µM colistin-treated BAM-OMVs. The experiment was performed one time. The selected image is representative for the entire EM grid. b SDS-PAGE of BAM-OMVs and purified BAM complex in DDM micelles unboiled (−) and boiled (+). The position of folded (BamAF) and unfolded (BamAU) is indicated, along with the lipoproteins BamB–BamE. The bands labelled with asterisk and X, correspond to truncated BamC and OmpX, respectively, as identified by mass spectrometry. The uncropped gel is provided as Source Data. The experiment was repeated independently 3 times with similar results. c Schematic representation of the assay setup comprising BAM-OMVs, colistin, chaperone-bound OmpT and QF peptide. d Progress curves for six different reaction mixes 1–6 as indicated. Data set is representative of n = 3 independent experiments with similar results. e Initial reaction velocities from D. n = 3 independent experiments. Data are presented as average values ± standard deviation. f Inhibitory effect of darobactin on BAM-mediated OmpT folding in OMVs. Data points indicate initial reaction velocities. The fitted curve corresponds to an IC50 value of 85 ± 7 nM. n = 2 independent experiments. Data are presented as average values ± standard deviation. The data underlying panels (d–f) are provided as Source Data.
