FIGURE 3.

Functional characterization of recombinant CGTs. A, C. difficile toxin B, C. sordellii lethal toxin, and C. novyi α-toxin were purified from B. megaterium and subjected to SDS-PAGE, and separated proteins were stained with Coomassie Blue. The bands indicated by arrowheads represent the respective purified toxins. B, in vitro glucosylation of Rac1, Ha-Ras, and RhoA with recombinant CGTs. C, intoxication of Vero cells with recombinant CGTs. Images were obtained after overnight (control (mock)), 1-h (100 pm toxin B), 6-h (1 nm lethal toxin), and overnight (1 nm α-toxin) treatment. D, InsP₆-induced in vitro cleavage of recombinant CGTs. Processing of CGTs (2 μg) was induced with increasing concentrations of InsP₆ as indicated and incubation for 1 h at 37 °C. E, processing of CGTs (2 μg) was induced with 100 μm InsP₆, followed by an incubation period as indicated at 37 °C.