FIGURE 4.

(A) Normalized hemolytic activity of wild type HlyA (left bar), single, triple and quadruple mutations within the secretion signal. The lysis of erythrocytes was quantified by measuring the release of hemoglobin by absorption spectroscopy at 544 nm. Control measurements shown to the right of the quadruple mutations lacked HlyA (acylated form), the acylase HlyC, Acyl-carrier protein (ACP), or a combination of these, in the assay. These results demonstrated that lysis was only induced in the presence of acylated HlyA. HlyA is as efficient in hemoglobin release as an SDS incubation (Thomas et al., 2014b) (not shown). Individual assays were performed in at three biological independent experiments and shown as scatter dot plots. (B) Summary of the secretion rates of wild type pro-HlyA (left), single, triple and quadruple mutations within the putative secretion signal. The value “WT” was taken from Lenders et al. (2016). Data represent the average of three biologically independent experiments and are shown as scatter dot plots.