Characterization of pilin protein membrane associations.
A. The cell wall, cytoplasmic and membrane fractions from wild-type (BD2528) cells were analysed by Western blotting with EF-G, ComEA, ComGA, ComGC and ComGG antisera.
B–E. Membranes isolated from cultures of (B, D) wild-type (BD2528) or (C, E) ComGD over-producing (BD5727) strains were incubated with buffers containing 0.75 M NaCl, 4 M urea, 8 M urea or 1% Triton X-100 as described in Experimental procedures. The soluble and insoluble fractions were separated by ultracentrifugation. Samples were immunoblotted with (B, D) ComGA, ComEA, ComGC, ComGG or (C, E) ComGD antisera.