FIG. 5.
KT interacts in vivo with TolA, TolB, TolQ, and TolR. Detergent-solubilized extracts of wild-type cells producing TolQHA and carrying the empty vector pIN-III-ompA2 (vector) or pKT (KT) producing the FLAG epitope-tagged colicin K N-terminal domain were subjected to immunoprecipitation using the anti-FLAG antibody (A). The total solubilized material (input [Inp]) and the precipitated material (IP) were loaded on a 12.5% acrylamide SDS-PAGE gel and immunodetected with the anti-HA and anti-FLAG monoclonal antibodies and with the anti-Pal, -TolA, -TolB, and -TolR polyclonal antibodies. Detergent-solubilized extracts of the same strains were subjected to immunoprecipitation using anti-TolA (B), anti-TolB (C), anti-TolR (D), and anti-HA (E) (precipitation of the HA epitope-tagged TolQ protein) antibodies. The corresponding mutant strain carrying the KT-producing vector has been used as an additional control for specificity. Immunodetected proteins are indicated on the right. Asterisks indicate heavy or light chains of immunoglobulins. Molecular mass markers (in kDa) are indicated on the left.