Molecular and enzymatic characterization of Schistosoma mansoni thioredoxin peroxidase.
The ability of Schistosoma mansoni to escape oxidative damage from immune system-generated reactive oxygen intermediates has been extensively documented. The limiting step in the parasite's detoxification process appears to be at the level of hydrogen peroxide neutralization. In the present study, the possible role of a novel class of antioxidant enzymes, thioredoxin peroxidase (TPx), in hydrogen peroxide neutralization by schistosomes was investigated. An expressed sequence tag was characterized from the Schistosoma Genome Initiative with high similarity to TPx from other organisms. The gene encodes a polypeptide containing 2 conserved active-site cysteines and flanking amino acids, and 60-70% identity with previously characterized TPx proteins. Recombinant schistosome TPx was enzymatically active and found to have thioredoxin-dependent hydrogen peroxide reducing activity of 4500 nmol hydrogen peroxide/min/mg protein. Native TPx activity was determined to be 48.1 nmol hydrogen peroxide/min/mg protein in adult worm homogenates compared with 46.9 for glutathione peroxidase. TPx activity was precipitated from adult worm homogenates with antibodies prepared against the recombinant protein. Western blotting with antibodies made against recombinant protein showed that TPx was expressed in both male and female adult worms. This is the first demonstration of a TPx activity in schistosomes and our results suggest that TPx plays a significant role in schistosome-host interactions.
Kwatia
M A
MA
Department of Biological Sciences, Illinois State University, Normal 61790-4120, USA.
Botkin
D J
DJ
Williams
D L
DL
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D018076:DNA, Complementary / Q000235:genetics; D020224:Expressed Sequence Tags; D005260:Female; D006861:Hydrogen Peroxide / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D010544:Peroxidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D054464:Peroxiredoxins; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000378:metabolism; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D009363:Neoplasm Proteins; D011994:Recombinant Proteins; D006861:Hydrogen Peroxide; D010544:Peroxidases; D054464:Peroxiredoxins