TCDB is operated by the Saier Lab Bioinformatics Group


1.C.5 The Channel-forming ε-toxin (ε-toxin) Family

The ε-toxin family consists of ε-toxin produced by Clostridium perfringens types B and D, which are responsible for a rapidly fatal enterotoxemia in sheep and other animals, as well as the mosquitocidal toxins, Mtx2 and Mtx3, produced by Bacillus sphaericus. These proteins are synthesized as relatively inactive prototoxins which are converted to active mature toxins by proteolytic removal of basic N-terminal peptides. ε-toxin increases intestinal and kidney cell permeability, forming a membrane complex of about 155 kDa which promotes efflux of intracellular K+ from target animal cells. The asymmetric pore complex (Nestorovich et al., 2010) is permeable to propidium ions and forms preferentially in the apical rather than the basolateral membrane (Petit et al., 2003). The mechanism of action of the mosquitocidal toxin is not known, but as these proteins are 20 to 27% identical to the ε-toxin of C. perfringens, they presumably function by a similar mechanism.

A 120-residue region of the ε-toxin of Clostridium perfringens (TC #1.C.5.1.1) shows significant sequence similarity to the pore-forming region of the pesticidal crystal protein Cry15Aa (insecticidal δ-endotoxin CryXVA(a)), and the first 77 residues of the epsilon toxin show sequence similarity with the first 82 residues of the beta-2 toxin of C. perfringens (TC #1.C.69). Cry15Aa is not demonstrably homologous to members of the channel-forming δ-endotoxin insecticidal crystal protein (ICP) family (TC #1.C.2). ε-toxin consists of a beta-barrel of 14 amphipatic beta strands (Popoff, 2011). The evidence presented by Knapp et al. (2009) suggests that the Aerolysin and RTX superfamilies may be distantly related.

ε-toxin (ETX) acts by heptamer formation and rapid permeabilization of target cell membranes for monovalent ions with later entry of calcium. Knapp et al. (2009) compared the primary structure of ETX with that of the channel-forming stretches of a variety of binding components of A-B-types of toxins such as Anthrax protective antigen (PA), C2II of C2-toxin and Ib of Iota-toxin and found homology to amino acids 151-180 of ETX. Site-directed mutagenesis of amino acids within the putative channel-forming domain resulted in changes of cytotoxicity and effects on channel characteristics in lipid bilayer experiments including changes in selectivity and partial channel block by methanethiosulfonate (MTS) reagents and antibodies against His(6)-tags from the trans-side of the lipid bilayer membranes.


The generalized transport reaction probably catalyzed by ε-toxin family members is:

small solutes (in) small solutes (out).


This family belongs to the: Aerolysin Superfamily.

References associated with 1.C.5 family:

Brown, K.L., and H.R. Whiteley. (1992). Molecular characterization of two novel crystal protein genes from Bacillus thuringiensis subsp. thompsoni. J, Bacteriol. 174: 549-557. 1729243
Chan, S.W., T. Thanabalu, B.Y. Wee, and A.G. Porter. (1996). Unusual amino acid determinants of host range in the Mtx2 family of mosquitocidal toxins. J. Biol. Chem. 271: 14183-14187. 8662969
Freedman, J.C., B.A. McClane, and F.A. Uzal. (2016). New insights into Clostridium perfringens epsilon toxin activation and action on the brain during enterotoxemia. Anaerobe 41: 27-31. 27321761
Knapp O., Maier E., Benz R., Geny B. and Popoff MR. (2009). Identification of the channel-forming domain of Clostridium perfringens Epsilon-toxin (ETX). Biochim Biophys Acta. 1788(12):2584-93. 19835840
Liu, J.W., A.G. Porter, B.Y. Wee, and T. Thanabalu. (1996). New gene from nine Bacillus sphaericus strains encoding highly conserved 35.8-kilodalton mosquitocidal toxins. Appl. Environ. Microbiol. 62: 2174-2176. 8787415
Miyata, S., J. Minami, E. Tamai, O. Matsushita, S. Shimamoto, and A. Okabe. (2002). Clostridium perfringens ε-toxin forms a heptameric pore within the detergent-insoluble microdomains of Madin-Darby canine kidney cells and rat synaptosomes. J. Biol. Chem. 277: 39463-39468. 12177068
Nestorovich, E.M., V.A. Karginov, and S.M. Bezrukov. (2010). Polymer partitioning and ion selectivity suggest asymmetrical shape for the membrane pore formed by epsilon toxin. Biophys. J. 99: 782-789. 20682255
Okumura, S., H. Saitoh, N. Wasano, H. Katayama, K. Higuchi, E. Mizuki, and K. Inouye. (2006). Efficient solubilization, activation, and purification of recombinant Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli. Protein Expr. Purif. 47: 144-151. 16307894
Petit, L., M. Gibert, D. Gillet, C. Laurent-Winter, P. Boquet, and M.R. Popoff. (1997). Clostridium perfringens ε-toxin acts on MDCK cells by forming a large membrane complex. J. Bacteriol. 179: 6480-6487. 9335299
Petit, L., M. Gilbert, A. Gourch, M. Bens, A. Vandewalle, and M.R. Popoff. (2003). Clostridium perfringens ε-toxin rapidly decreases membrane barrier permeability of polarized MDCK cells. Cell. Microbiol. 5: 155-164. 12614459
Popoff, M.R. (2011). Epsilon toxin: a fascinating pore-forming toxin. FEBS J. 278: 4602-4615. 21535407
Wioland, L., J.L. Dupont, F. Doussau, S. Gaillard, F. Heid, P. Isope, S. Pauillac, M.R. Popoff, J.L. Bossu, and B. Poulain. (2015). Epsilon toxin from Clostridium perfringens acts on oligodendrocytes without forming pores, and causes demyelination. Cell Microbiol 17: 369-388. 25287162