Plasminogen is recruited by Sbi and Efb.

<p>(A) Plasminogen bound to Sbi and Efb (lane 1 and 2). The accuracy of the recombinant staphylococcal proteins was confirmed as both, Sbi and Efb, bound to C3 (lane 3 and 4) and Sbi also bound IgG. Recombinant Sbi or Efb was separated by SDS-PAGE and either blotted to a membrane (lane 1–6) or silver stained (lane 7–8). Membranes were incubated with biotinylated PLG (PLG_b), C3, or HRP- coupled IgG and bound proteins were detected as indicated. (B) Binding of plasminogen to Sbi or Efb was confirmed by ELISA. Bacterial proteins (equimolar) were immobilized and PLG_b was applied and detected using streptavidin-HRP. Plasminogen binding to Sbi and Efb was comparable to plasminogen binding to the borrelial protein CRASP-5. Plasminogen did not bind to HSA or the plate (buffer). Data represent mean values ± standard deviations from three independent experiments. (C) Plasminogen was recruited to Sbi and Efb. Non-labeled plasminogen was applied to immobilized bacterial proteins. Bound proteins were eluted, separated by SDS-PAGE and plasminogen (92 kDa) was detected by Western blot analysis. CRASP-5 bound plasminogen in contrast to HSA and buffer. A representative experiment out of three is shown.