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J. Biol. Chem., Vol. 276, Issue 19, 15736-15740, May 11, 2001
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From Epsilon toxin is a potent toxin produced
by Clostridium perfringens types B and D, which are
responsible for a rapidly fatal enterotoxemia in animals. One of the
main properties of epsilon toxin is the production of edema. We have
previously found that epsilon toxin causes a rapid swelling of
Madin-Darby canine kidney cells and that the toxin does not
enter the cytosol and remains associated with the cell membrane by
forming a large complex (Petit, L., Gibert, M., Gillet, D.,
Laurent-Winter, C., Boquet, P., and Popoff, M. R. (1997) J. Bacteriol. 179, 6480-6487). Here, we report that epsilon toxin
induced in Madin-Darby canine kidney cells a rapid decrease of
intracellular K+, and an increase of Cl
Clostridium perfringens Epsilon Toxin Induces a Rapid
Change of Cell Membrane Permeability to Ions and Forms Channels in
Artificial Lipid Bilayers*
,,¶, and Centre National de Référence
Anaérobies, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris,
cedex 15, France and § Lehrstuhl für Biotechnologie,
Theodor-Boveri-Institut (Biozentrum) der Universität
Würzburg, Am Hubland, D-97074 Würzburg, Germany
and
Na+, whereas the increase of Ca2+ occurred
later. The entry of propidium iodide that was correlated with the loss
of cell viability monitored by the
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test
indicates that epsilon toxin formed large pores. In artificial lipid
bilayers, epsilon toxin caused current steps with a single-channel
conductance of 60 pS in 100 mM KCl, which represented
general diffusion pores. The channels were slightly selective for
anions, but cations could also penetrate. Epsilon toxin formed wide and
water-filled channels permeable to hydrophilic solutes up to a
molecular mass of at least 1 kDa, which probably represents the basic
mechanism of toxin action on target cells.
*
This study was supported in part by grants from the Deutsche
Forschungsgemeinschaft (Sonderforschungsbereich 487, Project A5) and
the Fonds der Chemischen Industrie and by a Caisse Autonome Nationale d'Assurance Maladíe fellowship (to L. P.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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