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  • Clostridium perfringens epsilon-toxin acts on MDCK cells by forming a large membrane complex.

Clostridium perfringens epsilon-toxin acts on MDCK cells by forming a large membrane complex.

Journal of bacteriology (1997-10-23)
L Petit, M Gibert, D Gillet, C Laurent-Winter, P Boquet, M R Popoff
ABSTRACT

Epsilon-toxin is produced by Clostridium perfringens types B and D and is responsible for a rapidly fatal enterotoxemia in animals, which is characterized by edema in several organs due to an increase in blood vessel permeability. The Madin-Darby canine kidney (MDCK) cell line has been found to be susceptible to epsilon-toxin (D. W. Payne, E. D. Williamson, H. Havard, N. Modi, and J. Brown, FEMS Microbiol. Lett. 116:161-168, 1994). Here we present evidence that epsilon-toxin cytotoxic activity is correlated with the formation of a large membrane complex (about 155 kDa) and efflux of intracellular K+ without entry of the toxin into the cytosol. Epsilon-toxin induced swelling, blebbing, and lysis of MDCK cells. Iodolabeled epsilon-toxin bound specifically to MDCK cell membranes at 4 and 37 labeled C and was associated with a large complex (about 155 kDa). The binding of epsilon-toxin to the cell surface was corroborated by immunofluorescence staining. The complex formed at 37 degrees C was more stable than that formed at 4 degrees C, since it was not dissociated by 5% sodium dodecyl sulfate and boiling.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Neuraminidase from Clostridium perfringens (C. welchii), Type VI, lyophilized powder, 6-15 units/mg protein (using 4MU-NANA), 2-10 units/mg protein (mucin)