Fig. 9.

EspP mutants that contain a lipid facing arginine near the middle of the β barrel inhibit Bam complex function. A and B. DPR959 transformed with pRLS5 or a pRLS5 derivative encoding the indicated EspP mutant were incubated at 37° C on LB agar plates containing ampicillin (100 μg/ml) and either no IPTG or 100 μM IPTG. C. AD202 transformed with pSCrhaB2 (vector), pJH207 (P_rhaB2-espPΔ5) or a pJH207 derivative encoding the indicated EspPΔ5 mutant were inoculated into LB containing trimethoprim (50 μg/ml) at OD₅₅₀=0.0001. The growth of each culture at 37° C was monitored at OD₅₅₀. D. AD202 transformed with pRLS5 and a derivative of pJH207 encoding wild-type EspPΔ5 or the indicated EspPΔ5 mutant were subject to pulse-chase labeling after the addition rhamnose and IPTG. Half of the cells were treated with PK, and immunoprecipitations were conducted using an antiserum generated against an EspP N-terminal peptide.