The Heterodimeric Protease Clostripain From Clostridium Histolyticum Is Encoded by a Single Gene

Mol Gen Genet. 1993 Jul;240(1):140-5. doi: 10.1007/BF00276893.


Clostripain (EC is a heterodimeric cysteine endopeptidase with strict specificity for Arg-Xaa peptidyl bonds. It is secreted by Clostridium histolyticum strains. For the first time we present evidence that both polypeptide chains of native clostripain are encoded by a single gene. DNA sequencing of two overlapping genomic DNA fragments revealed a single open reading frame (ORF) of 1581 nucleotides encoding a polypeptide of 526 amino acid residues. The ORF is preceded by canonical transcription signals and both chains of the clostripain heterodimer are completely represented by the deduced coding sequence. Most interestingly, the sequences coding for the light and the heavy chain are joined by a DNA stretch coding for a linker nonapeptide that is preceded by the C-terminal arginyl residue of the light chain and also ends with an arginyl residue. Heterologous expression of the gene in Escherichia coli yielded an enzyme capable of hydrolyzing the clostripain substrates N alpha-benzoyl-L-arginine ethyl ester (BAEE) and N-carbobenzoxy-L-arginine p-nitroanilide (Z-Arg-pNA).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Clostridium / enzymology*
  • Clostridium / genetics
  • Cysteine Endopeptidases / genetics*
  • Cysteine Endopeptidases / isolation & purification
  • DNA, Bacterial / genetics*
  • Escherichia coli
  • Gene Expression / genetics
  • Genes, Bacterial / genetics*
  • Molecular Sequence Data
  • Protein Sorting Signals / genetics
  • Transcription, Genetic / genetics


  • DNA, Bacterial
  • Protein Sorting Signals
  • Cysteine Endopeptidases
  • clostripain

Associated data

  • GENBANK/X63673