FIG. 5.

Identification of CspZ determinants required for the interaction with FH/FHL-1 and antibody elicited during infection. To identify the determinants of CspZ that are involved in the binding of CspZ to FH/FHL-1 and infection-induced antibody, a series of r-proteins was generated with site-directed mutations or as truncations. All proteins were generated using the B. burgdorferi B31MI-derived sequence. The full-length protein is labeled as Bb B31. Proteins with a decreased probability of coiled-coil formation either in cc1 or in cc2 were generated by site-directed mutagenesis and designated cc1m or cc2m, respectively. The truncations are designated by the amino acid regions that they span. (A) Equivalent loading was demonstrated using anti-His (α-His) and anti-CspZ (α-CspZ) antisera. The immunoreactivity of the proteins with anti-CspZ antibody that develops during infection of mice with B. burgdorferi B31MI was assessed by immunoblotting. FH binding was assessed using the ALBI (A) and ELISA (B) formats. Recombinant Hpk1 (BB0420) served as the negative control in the ELISA, and error bars represent standard deviation. All methods were as described in the text.