A recombinant thioredoxin-glutathione reductase from Fasciola hepatica induces a protective response in rabbits

Exp Parasitol. 2011 Dec;129(4):323-30. doi: 10.1016/j.exppara.2011.09.013. Epub 2011 Oct 1.

Abstract

Antioxidant systems are fundamental components of host-parasite interactions, and often play a key role in parasite survival. Here, we report the cloning, heterologous expression, and characterization of a thioredoxin glutathione reductase (TGR) from Fasciola hepatica. The deduced polypeptide sequence of the cloned open reading frame (ORF) confirmed the experimental N-terminus previously determined for a native F. hepatica TGR showing thioredoxin reductase (TR) activity. The sequence revealed the presence of a fusion between a glutaredoxin (Grx) and a TR domain, similar to that previously reported in Schistosoma mansoni and Echinococcus granulosus. The F. hepatica TGR sequence included an additional redox active center (ACUG; U being selenocysteine) located at the C-terminus. The addition of a recombinant selenocysteine insertion sequence (SECIS) element in the Escherichia coli expression vector, or the substitution of the native selenocysteine by a cysteine, indicated the relevance of this unusual amino acid residue for the activity of F. hepatica TGR. Rabbit vaccination with recombinant F. hepatica TGR reduced the worm burden by 96.7% following experimental infection, further supporting the relevance of TGR as a promising target for anti Fasciola treatments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • DNA, Helminth / chemistry
  • Fasciola hepatica / enzymology*
  • Fasciola hepatica / genetics
  • Fasciola hepatica / immunology
  • Fascioliasis / immunology*
  • Fascioliasis / prevention & control
  • Gene Expression Regulation, Enzymologic / immunology
  • Glutathione Reductase / chemistry
  • Glutathione Reductase / genetics
  • Glutathione Reductase / immunology*
  • Host-Parasite Interactions / immunology
  • Molecular Sequence Data
  • Protein Structure, Secondary
  • Rabbits
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Thioredoxin-Disulfide Reductase / chemistry
  • Thioredoxin-Disulfide Reductase / genetics
  • Thioredoxin-Disulfide Reductase / immunology*
  • Vaccination
  • Vaccines, Synthetic

Substances

  • DNA, Complementary
  • DNA, Helminth
  • Recombinant Proteins
  • Vaccines, Synthetic
  • Glutathione Reductase
  • Thioredoxin-Disulfide Reductase