Towards a Structural Comprehension of Bacterial Type VI Secretion Systems: Characterization of the TssJ-TssM Complex of an Escherichia coli Pathovar
Figure 2
TssJ interacts with the C-terminal domain of TssM.
(A) Solubilized extracts of E. coli K12 W3110 strain producing (+) or not (-) HA-tagged TssJ and FLAG-tagged TssM-ekto or -Nt or –Ct derivatives were subjected to immunoprecipitation with anti-FLAG-coupled beads. The total solubilized material (T) and the immunoprecipitated material (IP) were loaded on a 12.5%-acrylamide SDS PAGE, and immunodetected with anti-HA (TssJ; lower panel) and anti-FLAG (TssM-ekto and sub-domains; upper panel) monoclonal antibodies. Immunodetected proteins are indicated on the right. Molecular weight markers are indicated on the left. (B) Gel filtration showing the direct interaction of TssM-ekto with TssJ. The SDS-PAGE analysis of the fractions is shown on the left panel. The chromatogram of the gel filtration is shown on the right panel. (C) MALS/QELS/UV/RI analysis of the TssM-ekto/TssJ complex.