Fig 3.

Interactions between PrgJ and Pcf proteins. (A and C) GST affinity pulldowns. GST, GST-PcfC, GST-PcfF, or GST-PcfG produced in the E. coli strains listed in Table 1 were bound to glutathione beads and then mixed with PrgJ-His₆ (A) or PrgJ.K471E-His₆ (C) purified as described in the legend to Fig. 2. Following extensive washes, bound proteins were eluted from the beads with glutathione; proteins in E. coli cell lysates and eluates were identified by immunostaining with anti-GST and anti-His antibodies (for PrgJ-His₆). (B and D) Co²⁺ affinity pulldowns. Purified PrgJ-His₆ (B) or PrgJ.K471E-His₆ (D) was bound to Co²⁺ beads, and the protein-bead complexes were mixed with cell lysates containing the GST-tagged proteins shown. Following extensive washes, bound proteins were eluted from the beads with imidazole, and the proteins recovered in the eluates were identified by immunostaining with anti-GST and anti-His antibodies (for PrgJ-His₆).