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. 2021 Jul 8;10(7):830. doi: 10.3390/antibiotics10070830

Figure 3.

Figure 3

Characterization of AcrA mutants defective in TolC interaction. (a) Anti-AcrA Western blot analysis of samples prepared from BW25113 containing plasmids expressing the indicated AcrA mutant (diluted 4- folds). Sample prepared from plasmid-free BW25113 was also prepared and loaded without dilution (\) to serve as a control to highlight the difference in expression levels. (b) Anti-AcrB and Anti-AcrA Western blot analyses revealing the formation of disulfide bonded AcrA-AcrB complexes, which was reduced after incubation with BME. AcrA-P57C/AcrB-N191C, R128D (lane 1 and 3), AcrA-T217C/AcrB-S258C, R128D (lane 2 and 4). Molecular weight markers are labeled as “M” and the molecular weight of bands (kD) were indicated on the right. The expected bands for AcrA, AcrB, and disulfide bond linked AcrA-AcrB are marked on the left of the gels as A, B, and AB, respectively.