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 journalArticle

108 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
Publication statusPublished - 1997
Externally publishedYes

Fingerprint

Lethal Dose 50
Trypsin
Amino Acids
Peptides
Molecular Weight
Body Weight
Thermolysin
Clostridium perfringens
Metalloproteases
Polyacrylamide Gel Electrophoresis
Amino Acid Sequence
Mass Spectrometry
Lasers
Genes
trypsin drug combination chymotrypsin
caseinase

Keywords

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

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Microbiology (medical)
  • Microbiology

Cite this

Lambda-toxin of Clostridium perfringens activates the precursor of epsilon-toxin by releasing its N- and C-terminal peptides. / Minami, Junzaburo; Katayama, Seiichi; Matsushita, Osamu; Matsushita, Chieko; Okabe, Akinobu.

In: Microbiology and Immunology, Vol. 41, No. 7, 1997, p. 527-535.

Research output: Contribution to journalArticle

Minami, Junzaburo ; Katayama, Seiichi ; Matsushita, Osamu ; Matsushita, Chieko ; Okabe, Akinobu. / Lambda-toxin of Clostridium perfringens activates the precursor of epsilon-toxin by releasing its N- and C-terminal peptides. In: Microbiology and Immunology. 1997 ; Vol. 41, No. 7. pp. 527-535.
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