title: Clostridium perfringens iota toxin: binding studies and characterization of cell surface receptor by fluorescence-activated cytometry.
abstract: The binding characteristics of iota toxin, a binary enterotoxin produced by Clostridium perfringens type E, were studied by fluorescence-activated cytometry. 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. perfringens type E or Clostridium spiroforme culture supernatants, but not C. perfringens types C or D. Pretreatment of Vero cells with glycosidases or lectins did not affect Ib interactions, while pronase effectively prevented Ib binding to the cell surface. The Ib protomer (Ibp) bound to the cell surface, but trypsinization of Ibp was necessary for docking of the ADP-ribosylating component, iota a (Ia). Ia attached to cell-bound Ib within 10 min at 37 degrees C, but surface levels of Ia decreased 90% after 30 min and were undetectable 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.
authors.affiliation: Department of Immunology, U.S. Army Medical Research Institute of Infectious Diseases, Frederick, Maryland 21702-5011, USA. bradley.stiles@det.amedd.army.mil
authors.forename: B G
authors.initials: BG
authors.lastname: Stiles
authors.forename: M L
authors.initials: ML
authors.lastname: Hale
authors.forename: J C
authors.initials: JC
authors.lastname: Marvaud
authors.forename: M R
authors.initials: MR
authors.lastname: Popoff
mesh_terms: D036002:ADP Ribose Transferases; D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism* / Q000633:toxicity; D002460:Cell Line; D002522:Chlorocebus aethiops; D003016:Clostridium perfringens / Q000472:pathogenicity*; D004285:Dogs; D004768:Enterotoxins / Q000378:metabolism* / Q000633:toxicity; D006801:Humans; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014709:Vero Cells
chemical_list: D001427:Bacterial Toxins; D004768:Enterotoxins; D011956:Receptors, Cell Surface; C050737:iota toxin, Clostridium perfringens; D036002:ADP Ribose Transferases