FIG 5.

Only CtpA and its substrates bind to LbcA directly in vitro. Anti-FLAG M2 affinity resin was used to purify protein from E. coli lysates containing the empty pET-24b(+) vector (None) or a derivative encoding FLAG-LbcA. The purified anti-FLAG M2 agarose immunocomplexes were then incubated with approximately 1.5 μg of a purified prey protein (Prey) for 1 h, washed extensively in high-stringency buffer containing 0.5 M NaCl and 0.1% (wt/vol) Triton X-100, and analyzed by immunoblotting. Immunoblots are single representatives of results from several independent replicate experiments.