Extended Data Fig. 1: Determination of ATPase and PL transport activities of MlaFEDB.
From: Structural insights into outer membrane asymmetry maintenance in Gram-negative bacteria by MlaFEDB
a, Coomassie brilliant blue staining of the purified MlaFEDB on SDS-PAGE. b, The relative ATPase activities of purified MlaFEDB in liposomes or detergent. c, d, The relative ATPase activities of MlaFEDB in the presence of ADP (c) or AMP-PNP (d). e, f, SDS-PAGE analysis of MlaA-OmpF (e) or MlaFEDB (f) constitution ratio and orientation ratio in proteoliposomes of E. coli polar lipids or POPC with or without proteinase K treatment. g, FRET scan of PL transport assay using MlaA-OmpF wildtype or mutant MlaA(∆Asn41-Phe42)-OmpF in retrograde direction. h, Thin-layer chromatogram (TLC) showing transported PLs from IM or OM proteoliposomes to apo-MlaC. i, SDS-PAGE analysis of proteins involved in the transported system of Fig. g. j, FRET scan of PL transport assay containing IM proteoliposomes and apo-MlaC only in the presence or absence of ATP and MgCl2. k, l, FRET scan of PL transport assay using OM proteoliposomes containing lipid A in different ratios to PLs for anterograde (k) and retrograde direction (l). m, Loading control of lipids A and MlaA protein by western blot. Data in a, e-m are representative results from n = 3 independent experiments. Data in b-d, g, j-l represents mean ± s.d. (n = 3 independent experiments). Uncropped images for all gels are available online. Source data for panels b-d and g, j-l are available in Supplementary Data Set 1.
