Fine Mapping of the Interaction between C4b-Binding Protein and Outer Membrane Proteins LigA and LigB of Pathogenic Leptospira interrogans
Fig 2
Binding of recombinant proteins LcpA, LigAC, LigBC and whole L. interrogans to recombinant mutant C4BP molecules.
Microtiter plates were coated with LigAC (A), LigBC (B), LcpA (C) or LIC10301 (as negative control). After adding of each C4BP recombinant mutant or wild type (rec C4BP WT) protein (described in Fig 1B), binding was measured using rabbit polyclonal anti-human C4BP and peroxidase-conjugated anti-rabbit IgG. Each point represents the mean absorbance value at 492 nm +/- the SD of 3 independent experiments each performed in triplicate. The interaction of recombinant proteins of L. interrogans with rec C4BP WT was set as 100% binding. (D) Binding of C4BP mutant proteins to whole L. interrogans. Leptospires (1x108) were incubated with rec C4BP WT, C4BP mutants or PBS (negative control). To detect the C4BP binding to leptospires, polyclonal mouse anti-C4BP and FITC-conjugated anti-rabbit IgG were used. Each point represents the geometric mean fluorescence intensity (GMFI) +/- SE of 3 independent experiments each performed in triplicate. Data were analyzed using ANOVA test; *p<0.05; ***p<0.0001.