FIGURE 6.

α8 is required for dimerization as well as membrane insertion of TP0453. A, shown is the dimerization mode of TP0453 predicted by the PISA server. Monomers in TP0453 assemble side-by-side along an interface established by α8. B, shown are elution profiles of WT and mutant proteins on a Superdex^TM 75 HR column in 50 mm Tris (pH 7.5), 100 mm NaCl at 25 °C. The inset represents the calibration curve of the column. C, WT, α2^I62E, and α8^V249E were incubated in the absence and presence of LUVs under low and high salt conditions followed by glutaraldehyde cross-linking and SDS-PAGE/immunoblot analysis. Not shown are the results for α2^I62E, which were identical to WT. D, shown is BN-PAGE and immunoblot analysis of T. pallidum lysates (2 × 10⁸ organisms) solubilized in 2% n-dodecyl β-d-maltoside with 25 mm Tris (pH 7.0).