Fig. 6 | Nature Communications

Fig. 6

From: The complex of ferric-enterobactin with its transporter from Pseudomonas aeruginosa suggests a two-site model

Fig. 6

Effects of PfeA mutation in vivo. a Analysis of changes in the transcription of pfeA gene. PAO1 strain, its corresponding deletion mutant ∆pfeA and deletion mutants carrying the expression plasmid of PfeA wild-type (∆pfeA(pMMBpfeA) or the R480A-Q482A mutant (∆pfeA(pMMBpfeAR480AQ482A)) were grown in CAA medium supplemented with 10 µM enterobactin. The data were normalised relative to the reference gene uvrD, and are representative of three independent experiments performed in triplicate (n = 3). The results are given as a ratio between the values obtained for ∆pfeA, ∆pfeA(pMMBpfeA) and ∆pfeA(pMMBpfeAR480AQ482A) over those obtained for the PAO1 strain. b 55Fe-enterobactin binding at the bacterial surface. Cells were grown as in panel a, and were incubated for 15 min with 200 µM CCCP before initiation of transport assays by the addition of 500 nM 55Fe-enterobactin. After 30 min incubation, the radioactivity accumulated in the bacteria was counted. The results are expressed as pmol of 55Fe-enterobactin bound per ml of cells at an OD600 of 1. The experiments have been repeated three times. c 55Fe-enterobactin binding and uptake. Cells were grown as in panel a, and were incubated with or without 200 µM CCCP before initiation of transport assays by the addition of 500 nM 55Fe-enterobactin. After 30 min incubations, the radioactivity accumulated in the bacteria was counted. The results are expressed as pmol of 55Fe-enterobactin bound and transported per ml of cells at an OD600 of 1. The experiments have been repeated three times

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