FIGURE 1.

A, shown are structures of compounds used and visualization of the proteins from the pulldown assay. Structures of the inhibitory compounds, ME0052 and ME0055, and derivatives, ME0055-Aff and ME0052-Bio, used for the identification of target proteins are shown. Supplemental Protocol S1 details the syntheses. ME0055-Aff was used in affinity chromatography experiments to enrich for putative target proteins; ME0055-Bio was used for Far Western experiments. B, visualization of the protein fractions from the affinity chromatography procedure is shown. Lysates from E. coli O157 were incubated with ME0055-Aff and washed, and then proteins were eluted using 20 and 200 μm unlabeled ME0055. The beads were stripped using 1% acetic acid. Equal volumes of samples from each fraction were separated by SDS-PAGE and then visualized using Colloidal Blue Stain. Proteins were excised, subjected to in-gel trypsin digestion, and analyzed by LC-electrospray ionization tandem MS. Each protein identified from the gel is numbered sequentially and is listed in supplemental Table S4.