Lu/BCAM Adhesion Glycoprotein Is a Receptor for Escherichia coli Cytotoxic Necrotizing Factor 1 (CNF1) (original) (raw)
Figure 3
Cells not expressing Lu/BCAM do not respond to CNF1.
A) K562 leukemia cells, which do not express Lu/BCAM (top) and the isogenic cell line K562-Lu/BCAM expressing the receptor (bottom), were treated with GST-CNF1 for different time periods from 1 h to overnight (ON) as indicated. Uptake of the toxin was analyzed by the shift of modified RhoA in SDS-PAGE. B) GST-CNF1 was incubated with buffer or with recombinant BCAM in a molar ratio CNF1∶rBCAM of 1∶1, 1∶10 and 1∶100, respectively for 20 min. The mixture was added to K562-Lu/BCAM cells for 2 h. Cells were lysed and the deamidation of RhoA was analyzed by the shift of the modified GTPase in SDS-PAGE by Western-blotting. Data are representative for at least 3 independent experiments.