title: Inhibition of cytotoxicity by the Nhe cytotoxin of Bacillus cereus through the interaction of dodecyl maltoside with the NheB component.
abstract: Nhe ('nonhaemolytic enterotoxin') is a three-component cytotoxin implicated in the pathogenesis of diarrhoea by Bacillus cereus. Nhe forms pores in pure lipid bilayers, but the function of the individual components (NheA, NheB and NheC) remains unclear. NheB and NheC are structural homologues of ClyA, a pore-forming cytotoxin of Escherichia coli. The non-ionic detergent dodecyl maltoside (DDM) has been shown to inhibit haemolysis of ClyA. We used DDM as a probe to examine the response of the Nhe proteins to DDM micelles. At its critical micellar concentration (0.2 mM), DDM inhibited propidium uptake by the native Nhe complex in Vero and HT29 cell suspensions. Pre-incubation of NheC with DDM did not inhibit cytotoxicity. NheB exhibited marked changes in 1-anilinonaphthalene-8-sulphonic acid (ANS) fluorescence after pre-exposure to DDM. Pre-incubation of NheB with DDM resulted in large molecular weight complexes as detected by size exclusion chromatography and diffusion through sized dialysis membranes and prevented binding of NheB to Vero cell monolayers. These data support a model in which conformational changes and oligomerization of NheB are prerequisite events in the process of pore formation.
authors.affiliation: Department of Food Safety and Infection Biology, Norwegian School of Veterinary Science, Oslo, Norway.
authors.forename: Danh
authors.initials: D
authors.lastname: Phung
authors.forename: Per Einar
authors.initials: PE
authors.lastname: Granum
authors.forename: Richard
authors.initials: R
authors.lastname: Dietrich
authors.forename: Erwin
authors.initials: E
authors.lastname: Märtlbauer
authors.forename: Simon P
authors.initials: SP
authors.lastname: Hardy
mesh_terms: D000818:Animals; D001409:Bacillus cereus / Q000472:pathogenicity*; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D002460:Cell Line; D002850:Chromatography, Gel; D003603:Cytotoxins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000378:metabolism*; D005960:Glucosides / Q000378:metabolism*; D006801:Humans; D008970:Molecular Weight; D011485:Protein Binding; D011487:Protein Conformation; D055503:Protein Multimerization
chemical_list: D001427:Bacterial Toxins; D003603:Cytotoxins; D004791:Enzyme Inhibitors; D005960:Glucosides; C040358:dodecyl maltoside