Clostridium perfringens iota toxin: Binding studies and characterization of cell surface receptor by fluorescence-activated cytometry

The binding characteristics of iota toxin, a binary enterotoxin produced by Clostridium perfringens type E. were studied by fluorescence-activated cyt ometry. The proteolytically activated binding component of iota toxin, iota b (Ib), bound to various cell types when incubated at 4, 25, or 37 degrees C for 10 min. The binding of Ib was inhibited by antisera against C, perfr ingens type E or Clostridium spiroforme culture supernatants, but not C, pe rfringens types C or D, Pretreatment of Vero cells with glycosidases or lec tins did not affect Ib interactions, while pronase effectively prevented Ib binding to the cell surface. The Ib protomer (Ibp) bound to the cell surfa ce, but trypsinization of mp was necessary for docking of the ADP-ribosylat ing component, iota a (Ia), Ia attached to cell-bound Ib within 10 min at 3 7 degrees C, but surface levels of Ia decreased 90% after 30 min and were u ndetectable by 60 min. Detectable surface levels of Ib also diminished over time, and Western blot analysis suggested internalization or embedment of Ib into the membrane.