title: Changes in ganglioside content affect the binding of Clostridium perfringens epsilon-toxin to detergent-resistant membranes of Madin-Darby canine kidney cells.
abstract: Epsilon-toxin (ET) of Clostridium perfringens, which causes fatal enterotoxemia in ungulates, was previously shown to bind to and form a heptameric pore within the detergent-resistant membranes (DRMs) of MDCK cells. Depletion of cholesterol has also been shown to decrease the cytotoxicity of ET and its heptamerization. In this study, we investigated the effects of changes in sphingolipids, other DRM components of MDCK cells, on the cells' susceptibility to ET. Treatment with fumonisin B1 and PDMP, inhibitors of sphingolipid and glycosphingolipid syntheses, respectively, increased the susceptibility, while D609, a sphingomyelin synthesis inhibitor, had the opposite effect. The exogenous addition of ganglioside G(M1) dramatically decreased the ET binding, heptamerization and cytotoxicity. These effects were shown not to be due to ET binding to G(M1) or to denaturation of ET. We also found that the ET cytotoxicity towards MDCK cells decreased with an increase in culture time. In accordance with the resistance observed for prolonged cultured cells, G(M3), a major ganglioside component, increased and sialidase treatment increased their susceptibility. These results suggest that membrane-anchored sialic acid of G(M3) within DRMs inhibits ET binding, leading to prevention of the heptamerization of ET and cell death. It is also suggested that sialidase produced by this organism aids the targeting of ET to MDCK cells.
authors.affiliation: Department of Microbiology, Faculty of Medicine, Kagawa University, Kita-gun, Japan.
authors.forename: Seiko
authors.initials: S
authors.lastname: Shimamoto
authors.forename: Eiji
authors.initials: E
authors.lastname: Tamai
authors.forename: Osamu
authors.initials: O
authors.lastname: Matsushita
authors.forename: Junzaburo
authors.initials: J
authors.lastname: Minami
authors.forename: Akinobu
authors.initials: A
authors.lastname: Okabe
authors.forename: Shigeru
authors.initials: S
authors.lastname: Miyata
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism* / Q000633:toxicity; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003016:Clostridium perfringens / Q000378:metabolism* / Q000472:pathogenicity*; D003902:Detergents; D004285:Dogs; D037341:Fumonisins / Q000494:pharmacology; D005677:G(M1) Ganglioside / Q000494:pharmacology; D005679:G(M3) Ganglioside / Q000378:metabolism; D005732:Gangliosides / Q000378:metabolism*; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism; D009025:Morpholines / Q000494:pharmacology; D009439:Neuraminidase / Q000494:pharmacology
chemical_list: D001427:Bacterial Toxins; C039811:Clostridium perfringens epsilon-toxin; D003902:Detergents; D037341:Fumonisins; D005679:G(M3) Ganglioside; D005732:Gangliosides; D008563:Membrane Lipids; D009025:Morpholines; D005677:G(M1) Ganglioside; C056933:fumonisin B1; C033110:RV 538; D009439:Neuraminidase