Fasciola hepatica: Heterologous Expression and Functional Characterization of a Thioredoxin Peroxidase

• Published in: Experimental parasitology Vol. 95; no. 1; pp. 63 - 70
• Main Authors: Salazar-Calderón, Mara, Martín-Alonso, José M., Ruiz de Eguino, Arantxa D., Casais, Rosa, Marín, M.Soledad, Parra, Francisco
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.05.2000; Elsevier
• Subjects: Amino Acid Sequence; Animals; antioxidant protein; Antioxidants - chemistry; Antioxidants - isolation & purification; Antioxidants - metabolism; Base Sequence; Biological and medical sciences; Blotting, Northern; Blotting, Southern; Blotting, Western; Cattle; Cloning, Molecular; DNA, Complementary - chemistry; DNA, Helminth - chemistry; Electrophoresis, Polyacrylamide Gel; excretory–secretory extract (ESE); Fasciola hepatica; Fasciola hepatica - enzymology; Fasciola hepatica - genetics; Fe3+/O2/thiol metal-catalyzed oxidation systems (MCO); Fundamental and applied biological sciences. Psychology; Gene Expression; glutamine synthase; glutamine synthetase (GS); Host parasite relation; pathogenicity; Invertebrates; Molecular Sequence Data; Nemathelminthia. Plathelmintha; Neoplasm Proteins; Open Reading Frames; Peroxidases - chemistry; Peroxidases - genetics; Peroxidases - metabolism; Peroxiredoxins; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; RNA, Messenger - analysis; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Thioredoxin peroxidase; thioredoxin peroxidase (TPx); TSA protein; glutamine synthetase (GS); Fasciola hepatica; TSA protein; Fe3+/O2/thiol metal-catalyzed oxidation systems (MCO); antioxidant protein; thioredoxin peroxidase (TPx); excretory–secretory extract (ESE); Host parasite relation; Nucleotide sequence; Enzyme; Plathelmintha; Parasite; Antioxidant; Gene expression; Thioredoxin; Trematoda; Biological activity; Peroxidases; Complementary DNA; Helmintha; Peroxidase; Oxidoreductases; Recombinant protein; Exocrine secretion; Invertebrata; Heterologous; Aminoacid sequence
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1006/expr.2000.4495

Salazar-Calderón, M., Martín-Alonso, J. M., Ruiz de Eguino, A. D., Casais, R., Marín, M. S., and Parra, F. 2000. Fasciola hepatica: Heterologous expression and functional characterization of a thioredoxin peroxidase. Experimental Parasitology95, 63–70. A Fasciola hepatica cDNA clone of 779 bp was isolated from an adult worm cDNA expression library by immunological screening using a rabbit serum against the excretory–secretory antigens. The nucleotide sequence of the cDNA revealed the presence of an open reading frame of 582 bp which encoded a 194-amino-acid-residue polypeptide (Mr 21,723 Da) showing a high degree of homology to thioredoxin peroxidases. This putative antioxidant protein gene was expressed in Escherichia coli as a GST fusion protein. The recombinant fusion protein showed in vitro antioxidant properties and protected rabbit muscle enolase and E. coli glutamine synthetase from inactivation by nonenzymatic Fe3+/O2/DTT or Fe3+/O2/ascorbate metal-catalyzed oxidation systems.