Figure 2

Paracoccidioides FBA is a plasminogen-binding protein. (A) SDS-PAGE analysis of the rFBA of Paracoccidioides. The recombinant protein was obtained by heterologous expression in E. coli. The bacteria total protein extract, before (lane 1), and after (lane 2) the induction with IPTG; the recombinant protein fused to gluthatione S-transferase (GST) (lane 3) and the purified rFBA (lane 4). (B) Far-western analysis. Increasing concentrations of the rFBA (0.1 to 3.0 μg) were fractionated by SDS-PAGE (12%) and transfered to a nitrocellulose membrane that was subsequently incubated with hPLg, anti-hPlg antibodies produced in mouse and anti-mouse immunoglobulin G (IgG) coupled to alkaline phosphatase. The reaction was developed using 5-Bromo-4-chloro-3-indolyl phosphate (BCIP) and Nitro Blue Tetrazolium (NBT).