Figure 5. Binding specificity of soluble CL-12 towards properdin variants.
(A) Illustrative diagram of monomer-like properdin complex. (B) SEC analysis of properdin. Inset: SDS-PAGE analysis of properdin under reducing condition. (C) A. fumigatus were incubated with fP-Dpl (10%) restored with purified serum properdin (10 µg/ml) or properdin variants (fP T: properdin tetramer; fP DT: properdin dimer/trimer; fP M: monomer-like properdin, 10 µg/ml). C3b deposition were analyzed and expressed as % Positive events with respect to the fP-Dpl alone with mean ± S.E.M from three independent experiments. (D) sCL-12 (5 µg/ml) was preincubated with the fungus prior to addition of the serum treated with Cp40 (6 µM) plus the properdin variants. Binding of properdin variants was analyzed in sCL-12-positive region (R1 in FSC vs SSC of the fungus) and expressed as % properdin/CL-12-double positive events with mean ± S.E.M from three independent experiments. (E) Binding of properdin tetramer (20 µg/ml) was determined on A. niger as above. Results are representative of at least six independent experiments.
Figure 5—figure supplement 1. Binding of properdin variants to A.niger in the presence of soluble CL-12.
