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Infection and Immunity, July 2000, p. 3848-3853, Vol. 68, No. 7
Unité des Toxines Microbiennes,
Institut Pasteur, 75724 Paris Cedex 15, France,1
and Department of Microbiology, Faculty of Medicine, Kagawa
Medical University, Kita-gun, Kagawa, 761-0793 Japan2
Received 29 November 1999/Returned for modification 21 January
2000/Accepted 1 April 2000
Iota-toxin is produced by Clostridium perfringens type
E strains and consists of two independent components, the enzymatic and
binding components, referred to as Ia and Ib, respectively. A
recombinant C. perfringens strain, strain 667/pMRP147,
produced processed Ia and partially processed Ib, while a recombinant
C. perfringens type A strain, strain TS133/pMRP147, in
which the VirR-VirS two-component system is inactivated, produced only
precursor forms of Ia and Ib. This suggests that iota-toxin is
processed by a VirR-VirS-responsive protease, although not completely
in the recombinant type A strain. The precursor forms of Ia and Ib were
purified from cultures of the latter strain, and their proteolytic activation was examined. Treatment with proteases cleaved off small
peptides (9 to 13 amino acid residues) and a 20-kDa peptide from the N
termini of the Ia and Ib precursors, respectively, leading to their
active forms. They were activated efficiently by
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Copyright © 2000, American Society for Microbiology. All rights reserved.
Clostridium perfringens Iota-Toxin
Requires Activation of Both Binding and Enzymatic Components for
Cytopathic Activity
-chymotrypsin,
pepsin, proteinase K, subtilisin, and thermolysin but only weakly by
trypsin, as demonstrated by the cell-rounding assay.
-Protease from
the C. perfringens type E strain, which was found to be a
zinc-dependent protease related to thermolysin, activated iota-toxin as
efficiently as did
-chymotrypsin. These results suggest that
-protease is most responsible for the activation of iota-toxin in
type E strains.
*
Corresponding author. Mailing address: Unité des
Toxines Microbiennes, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris
Cedex 15, France. Phone: 33 1 45 68 83 07. Fax: 33 1 40 61 31 23. E-mail: mpopoff{at}pasteur.fr.
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