Figure 5

Localization of the binding sites involved in the interaction of IsdB with vWF. (A) Concentration dependent-binding of IsdB NEAT1-NEAT2 to immobilized vWF or the vWF A1 domain. vWF and the recombinant A1 domain were immobilized onto microtiter wells and incubated with increasing concentrations of IsdB NEAT1-NEAT2. Complex formation was detected by addition to the wells of a polyclonal rabbit anti-IsdB IgG followed by HRP-conjugated goat anti-rabbit IgG. (B) Binding of the A1 domain to IsdB proteins. IsdB NEAT1–NEAT2 protein and its NEAT1 and NEAT2 subregions were immobilized onto microtiter wells and incubated with increasing concentrations of the A1 domain. Ligand binding was detected as reported in Fig. 1C. The data points are the means ± SD from three independent experiments, each performed in duplicate.