Fig 6.

Biochemical analysis of protein–protein interactions involving pilus proteins. A. SDS-PAGE analysis of recombinant pilus proteins rAP1_M6, rBP_M6, rAP1_M1 and rAP1_M3; lane 2 contains a main band of 62 kDa corresponding to rBP_M6 and a secondary band of about 90 kDa, which corresponds to an impurity from E. coli. B. Far Western blot analysis using biotinylated rAP1_M6 (rAP1_M6*) as probe against the recombinant proteins shown in (A); Far Western blot assay of biotinylated rBP_M6*, rAP1_M1* and rAP1_M3* as probes against the respective pilus proteins loaded on SDS-PAGE. C. ELISA saturation kinetic analysis of biotinylated rAP1_M6* protein bound to immobilized unlabelled rAP1_M6. All experiments were conducted in triplicate with reproducible results, and mean and standard deviation values are reported.