Figure 4.

Proteolysis of TolC derivatives on the extracellular surface of whole cells. Cleavage of TolC carrying TEV protease sites was monitored by Western blotting. TEV protease was added to intact cells for 3 hr at 30°C. After inactivation of TEV protease, cells were suspended in sample buffer and subjected to SDS/PAGE. wt, wild-type TolC. Numbers indicate the amino acid of TolC after which a TEV protease site was inserted. −, no TEV protease added to whole cells. +, TEV protease (70 μg/ml final concentration) added to whole cells.