Interaction of Clostridium perfringens epsilon-toxin with biological and model membranes: A putative protein receptor in cells

Biochim Biophys Acta. 2015 Mar;1848(3):797-804. doi: 10.1016/j.bbamem.2014.11.028. Epub 2014 Dec 5.

Abstract

Epsilon-toxin (ETX) is a powerful toxin produced by some strains of Clostridium perfringens (classified as types B and D) that is responsible for enterotoxemia in animals. ETX forms pores through the plasma membrane of eukaryotic cells, consisting of a β-barrel of 14 amphipathic β-strands. ETX shows a high specificity for certain cell lines, of which Madin-Darby canine kidney (MDCK) is the first sensitive cell line identified and the most studied one. The aim of this study was to establish the role of lipids in the toxicity caused by ETX and the correlation of its activity in model and biological membranes. In MDCK cells, using cell counting and confocal microscopy, we have observed that the toxin causes cell death mediated by toxin binding to plasma membrane. Moreover, ETX binds and permeabilizes the membranes of giant plasma membrane vesicles (GPMV). However, little effect is observed on protein-free vesicles. The data suggest the essential role of a protein receptor for the toxin in cell membranes.

Keywords: Epsilon-toxin; GUV; Giant plasma membrane vesicles; MDCK cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Bacterial Toxins / genetics
  • Bacterial Toxins / metabolism*
  • Bacterial Toxins / pharmacology
  • Cell Membrane / metabolism*
  • Cell Survival / drug effects
  • Clostridium perfringens / metabolism
  • Dogs
  • Flow Cytometry
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Madin Darby Canine Kidney Cells
  • Membrane Lipids / metabolism
  • Membrane Proteins / metabolism*
  • Microscopy, Confocal
  • Protein Binding
  • Receptors, Cell Surface / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / pharmacology
  • Unilamellar Liposomes / metabolism

Substances

  • Bacterial Toxins
  • Clostridium perfringens epsilon-toxin
  • Membrane Lipids
  • Membrane Proteins
  • Receptors, Cell Surface
  • Recombinant Fusion Proteins
  • Unilamellar Liposomes
  • Green Fluorescent Proteins