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Biochim Biophys Acta. 2016 Nov;1858(11):2681-2688. doi: 10.1016/j.bbamem.2016.07.009. Epub 2016 Jul 22.

Oligomer formation of Clostridium perfringens epsilon-toxin is induced by activation of neutral sphingomyelinase.

Author information

1
Department of Microbiology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Yamashiro-cho 180, Tokushima 770-8514, Japan.
2
Division of Microbiology and Infectious Diseases, Niigata University Graduate School of Medical and Dental Sciences, 2-5274, Gakkocho-dori, Chuo-ku, Niigata 951-8514, Japan.
3
Department of Microbiology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Yamashiro-cho 180, Tokushima 770-8514, Japan. Electronic address: nagahama@ph.bunri-u.ac.jp.

Abstract

BACKGROUND:

Clostridium perfringens epsilon-toxin is responsible for fatal enterotoxemia in ungulates. The toxin forms a heptamer in the lipid rafts of Madin-Darby Canine Kidney (MDCK) cells, leading to cell death. Here, we showed that epsilon-toxin requires neutral sphingomyelinase (nSMase) activity during oligomerization.

METHODS:

We tested the role of nSMase in the oligomerization of epsilon-toxin using specific inhibitors, knockdown of nSMase, formation of ceramide, and localization of epsilon-toxin and ceramide by immunofluorescence staining.

RESULTS:

Epsilon-toxin induced the production of ceramide is a dose- and time-dependent manner in ACHN cells. GW4869, an inhibitor of nSMase, inhibited ceramide production induced by the toxin. GW4869 and knockdown of nSMase blocked toxin-induced cell death and oligomer formation of epsilon-toxin. Confocal microscopy images showed that the toxin induced ceramide clustering and colocalized with ceramide.

CONCLUSIONS:

These results demonstrated that oligomer formation of epsilon-toxin is facilitated by the production of ceramide through activation of nSMase caused by the toxin.

GENERAL SIGNIFICANCE:

Inhibitors of nSMase may confer protection against infection.

KEYWORDS:

C. perfringens epsilon-toxin; Ceramide; Neutral sphingomyelinase; Oligomer

PMID:
27453200
DOI:
10.1016/j.bbamem.2016.07.009
[Indexed for MEDLINE]
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