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. 1987 Jul;55(7):1610–1615. doi: 10.1128/iai.55.7.1610-1615.1987

Biochemical studies on the effect of Clostridium difficile toxin B on actin in vivo and in vitro.

M J Mitchell, B E Laughon, S Lin
PMCID: PMC260566  PMID: 3596804

Abstract

We describe a simplified procedure for purification of Clostridium difficile toxin B. In this procedure, cytotoxicity is associated with a single protein band with a molecular mass of 230 kilodaltons. We used direct fluorescent staining of actin filaments to study the effect of this toxin on cultured cells. Morphologic changes were preceded by a decrease in the number and length of stress fibers followed by their disappearance with condensation of cellular actin around the nucleus. We then showed that cells treated with either cytochalasin B or toxin B had a significant increase in the monomeric actin pool as quantitated by DNase I inhibition. In contrast to the cytochalasins, toxin B had no direct effect on the rate or extent of actin polymerization or network formation in vitro. Cytoplasmic extracts of toxin B-treated cells had a significantly lower level of modulating activity on actin assembly and interactions in vitro compared with extracts of untreated cells. These results suggest that the action of toxin B on cells is due to direct or indirect effects on cellular proteins involved in controlling the state of actin assembly in the cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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