Characterization of two cysteine proteases secreted by Blastocystis ST7, a human intestinal parasite

Parasitol Int. 2012 Sep;61(3):437-42. doi: 10.1016/j.parint.2012.02.007. Epub 2012 Feb 25.

Abstract

Blastocystis spp. are unicellular anaerobic intestinal parasites of both humans and animals and the most prevalent ones found in human stool samples. Their association with various gastrointestinal disorders raises the questions of its pathogenicity and of the molecular mechanisms involved. Since secreted proteases are well-known to be implicated in intestinal parasite virulence, we intended to determine whether Blastocystis spp. possess such pathogenic factors. In silico analysis of the Blastocystis subtype 7 (ST7) genome sequence highlighted 22 genes coding proteases which were predicted to be secreted. We characterized the proteolytic activities in the secretory products of Blastocystis ST7 using specific protease inhibitors. Two cysteine proteases, a cathepsin B and a legumain, were identified in the parasite culture supernatant by gelatin zymographic SDS-PAGE gel and MS/MS analysis. These proteases might act on intestinal cells and disturb gut function. This work provides serious molecular candidates to link Blastocystis spp. and intestinal disorders.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Blastocystis / cytology
  • Blastocystis / enzymology*
  • Blastocystis / genetics*
  • Cathepsin B / genetics
  • Cathepsin B / isolation & purification
  • Cathepsin B / metabolism
  • Cysteine Endopeptidases / genetics
  • Cysteine Endopeptidases / isolation & purification
  • Cysteine Endopeptidases / metabolism
  • Cysteine Proteases / genetics
  • Cysteine Proteases / isolation & purification
  • Cysteine Proteases / metabolism*
  • DNA, Protozoan / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Intestines / parasitology
  • Molecular Sequence Data
  • Protease Inhibitors / metabolism
  • Proteomics
  • Tandem Mass Spectrometry
  • Virulence Factors / genetics
  • Virulence Factors / isolation & purification
  • Virulence Factors / metabolism

Substances

  • DNA, Protozoan
  • Protease Inhibitors
  • Virulence Factors
  • Cysteine Proteases
  • Cysteine Endopeptidases
  • Cathepsin B
  • asparaginylendopeptidase