Binding of human plasminogen (plg) to recombinant proteins rPDHA-D.

<p>(A) Western blot analysis of immobilized total proteins of <i>M</i>. <i>pneumoniae</i> M129 and recombinant proteins rPDHA-D incubated with human plasminogen. Bound plasminogen was detected with rabbit anti-plasminogen and anti-rabbit IgG. Treatment of blotted BSA (66.5 kDa) served as negative control. (B) ELISA experiments to characterize the binding of different concentrations of plasminogen to immobilized rPDHA-D. Detection of plasminogen was carried out as described. Data represent means and standard deviations of eight parallels. (C) Influence of pre-incubation of rPDHA-D with the corresponding antisera on the interaction of human plasminogen with recombinant proteins. Plasminogen was pre-incubated with the sera against the subunits and added to immobilized recombinant proteins. Detection of bound plasminogen in comparison with controls using pre-immune serum (PIS) for pre-incubation was carried out by using anti-plasminogen and peroxidase-conjugated anti-rabbit IgG. Data represent means and standard deviations of eight parallels (*** = P<0.001; student’s t-test). (D) Concentration-dependent binding of plasminogen pre-incubated with the antiserum to whole proteins and with the mixture of anti-rPDHA-D to immobilized total proteins of <i>M</i>. <i>pneumoniae</i> M129 (M.p.). Plasminogen incubated with guinea pig pre-immune serum (PIS) was used as control. Means and standard deviations of eight parallels are shown (** = P<0.01; student’s t-test).