Interaction of cholera toxin and Escherichia coli enterotoxin with isolated intestinal epithelial cells.

The interaction of biologically active 125I-labeled cholera toxin with isolated chick intestinal epithelial cells involves a large number (approx 1.7 10(6)/cell) of high-affinity (Kd = 8-9 X 10(-9) M) binding sites that belong to a single class. Binding of iodotoxin to the cells occurs rapidly, is half-maximal within 1 min, and is complete in 3-7 min (at 37 degrees C) depending on the toxin concentration. Toxin binding is saturable and includes only a small contribution from nonspecific sites. Ligand competition studies suggest that the isolated B subunit of choleragen (CT-B) behaves in an almost identical fashion to the holotoxin (CT), whereas the A subunit shows no detectable activity in competitive binding. Assays for cAMP indicate that neither that A nor the B subunits of CT contain any activity for increasing the level of intracellular cAMP. B subunit, when incubated with CT, inhibits CT-induced elevation of cAMP in a dose-dependent manner. Preincubation of 125I-CT with various concentrations of ganglioside GM1 also shows a dose-dependent inhibitory effect on the binding activity of the toxin. Pretreatment of CT with increasing concentrations of GM1 results in a progressive decrease in toxin-induced formation of cAMP. Escherichia coli heat-labile enterotoxin, which is known to alter intestinal function via a mechanism similar to that of CT, has binding and biological effects very similar to those of CT.