title: A recombinant thioredoxin-glutathione reductase from Fasciola hepatica induces a protective response in rabbits.
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.
authors: Departamento de Bioquímica y Biología Molecular, Instituto Universitario de Biotecnología de Asturias, Universidad de Oviedo, 33006 Oviedo, Spain.
authors: Gabriela
authors: G
authors: Maggioli
authors: Fernando
authors: F
authors: Silveira
authors: José M
authors: JM
authors: Martín-Alonso
authors: Gustavo
authors: G
authors: Salinas
authors: Carlos
authors: C
authors: Carmona
authors: Francisco
authors: F
authors: Parra
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry; D005210:Fasciola hepatica / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D005211:Fascioliasis / Q000276:immunology* / Q000517:prevention & control; D015971:Gene Expression Regulation, Enzymologic / Q000276:immunology; D005980:Glutathione Reductase / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006790:Host-Parasite Interactions / Q000276:immunology; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013880:Thioredoxin-Disulfide Reductase / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D014611:Vaccination; D014614:Vaccines, Synthetic
chemical_list: D018076:DNA, Complementary; D018509:DNA, Helminth; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D005980:Glutathione Reductase; D013880:Thioredoxin-Disulfide Reductase