Lambda-toxin of Clostridium perfringens activates the precursor of epsilon-toxin by releasing its N- and C-terminal peptides

Junzaburo Minami, Seiichi Katayama, Osamu Matsushita, Chieko Matsushita, Akinobu Okabe

Research output: Contribution to journalArticlepeer-review

112 Citations (Scopus)

Abstract

The effect of λ-toxin, a thermolysin-like metalloprotease of Clostridium perfringens, on the inactive ε-prototoxin produced by the same organism was examined. When the purified ε-prototoxin was incubated with the purified ε-toxin at 37 C for 2 hr, the 32.5-kDa ε-prototoxin was processed into a 30.5-kDa polypeptide, as determined by SDS-polyacrylamide gel electrophoresis. A mouse lethality test showed that the treatment activated the prototoxin: the 50% lethal doses (LD50) of the prototoxin with and without λ-toxin treatment were 110 and 70,000 ng/kg of body weight, respectively. The lethal activity of the prototoxin activated by λ-toxin was comparable to that with trypsin plus chymotrypsin and higher than that with trypsin alone: LD50 of the prototoxin treated with trypsin and trypsin plus chymotrypsin were 320 and 65 ng/kg of body weight, respectively. The ε- toxin gene was cloned and sequenced. Determination of the N-terminal amino acid sequence of each activated ε-prototoxin revealed that λ-toxin cleaved between the 10th and 11th amino acid residues from the N-terminus of the prototoxin, while trypsin and trypsin plus chymotrypsin cleaved between the 13th and 14th amino acid residues. The molecular weight of each activated ε- prototoxin was also determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The C-terminus deduced from the molecular weight is located at the 23rd or 30th amino acid residue from the C-terminus of the prototoxin, suggesting that removal of not only N-terminal but also C- terminal peptide is responsible for activation of the prototoxin.

Original languageEnglish
Pages (from-to)527-535
Number of pages9
JournalMICROBIOLOGY and IMMUNOLOGY
Volume41
Issue number7
DOIs
Publication statusPublished - Jan 1 1997
Externally publishedYes

Keywords

  • Activation of prototoxin
  • Clostridium perfringens
  • Protease
  • Toxin
  • Virulence

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

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