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. 2017 May 2;8:747. doi: 10.3389/fmicb.2017.00747

Figure 3.

Figure 3

Activity and processing of constitutively active IutY point mutants. (A) Schematic representation of the P. putida IutY protein drawn as in Figure 2A. The position of the IutY single mutations tested is indicated. (B) β-Galactosidase activity of the P. putida ΔiutY mutant strain bearing the transcriptional fusion iutA::lacZ (pMPK4 plasmid) and the pMMB67EH (empty, -), pMMBK1-HA (WT), pMMBK1-HA-D210, -R231Q, -A240E, -F251S, -V253D, -W301G, or -LtoP construct expressing the corresponding IutY protein variant grown in low iron medium supplemented with 1 mM IPTG in the absence (light gray bars) or presence (dark gray bars) of aerobactin. (C) The P. putida ΔiutY strain expressing the indicated N-terminally HA-tagged IutY mutant protein was grown in the absence (–) or presence (+) of aerobactin. Proteins were detected by Western-blot using an anti-HA antibody. Position of the protein fragments (full length IutY and σIutY domain) and the molecular size marker (in kDa) are indicated.