FIG. 5.

The intramolecular coiled-coil motif in LcrV is not required for LcrV's dimerization. E. coli (Novablue) cells carrying pBAD18 (empty vector; lane 1), pAraV18 (full-length LcrV; lane 2), pMH61 (LcrVΔα-7; lane 3), pMH13 (LcrVΔα-12, lane 4), and pMH122 (LcrV311, lane 5) were grown in LB plus arabinose for 3 h. Cells were harvested by centrifugation, and cellular fractions were precipitated with trichloroacetic acid overnight at 4°C. Samples were separated by SDS-PAGE in a 15% polyacrylamide gel and analyzed by immunoblotting with anti-LcrV. Positions of the LcrV dimers are indicated by arrowheads.