FIG. 3.
Analysis of the complement regulator-binding capabilities of native and recombinant CRASP-1 molecules of B. burgdorferi sensu lato strains. Binding capabilities of FHL-1 and/or factor H to cell lysates or to recombinant CRASP-1 molecules were analyzed by the ligand affinity blot technique. Whole-cell lysates of strains ZS7, MMS, and ZQ1 (A) or recombinant CRASP-1Bb, CRASP-1Ba, CRASP-1Bg, CRASP-1βBg, or CRASP-3Bb (400 ng/lane) (B) were subjected to 10% Tris-Tricine-SDS-PAGE and blotted to nitrocellulose membranes. These membranes were then incubated with either recombinant FHL-1 or with NHS for detection of factor H binding. Bound proteins were visualized using antiserum (αSCR1-4) specific for the N-terminal region of FHL-1 or MAb VIG8 specific for the C-terminal region of factor H. Flagellin and Hsp70 were detected using MAbs LA22.1 or LA3, respectively. The mobilities of the marker proteins are indicated to the right.