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Flow cytometry of E. coli UL3 fhuA transformed with plasmids pT7-6 (vector without fhuA) (+), pfhuA8 [FhuA (wild type)] (•), pfhuA5 [FhuA(C329S) Cys-318] (∗), pfhuA4 [FhuA(C318S) Cys-329] (⧫), pfhuA7 [FhuA(V347C)] (▪), and pfhuA6 [FhuA(D336C)] (▴) after labeling with 0.5 mM F-M for 0.5, 1.5, 5, or 15 min at 30°C. The reaction was stopped by adding 20 mM DTT. The fluorescence values showed a standard deviation of 10%.
