title: Cloning and characterization of Plasmodium vivax thioredoxin peroxidase-1.
abstract: Reactive oxygen species produced from hemoglobin digestion and the host immune system could have adverse effects on malaria parasites. To protect themselves, malaria parasites are highly dependent on the antioxidant enzymes, including superoxide dismutases and thioredoxin-dependent peroxidases. To date, several thioredoxin peroxidases (TPx) have been characterized in Plasmodium falciparum, but the TPx in Plasmodium vivax has not yet been characterized. The complete sequence of gene coding for thioredoxin peroxidase-1 of P. vivax (PvTPx-1) was amplified by PCR and cloned. Using the recombinant PvTPx-1 (rPvTPx-1), polyclonal antibody was produced in mice for immunolocalization of the enzyme in the parasite. The antioxidant activity of rPvTPx-1 was evaluated by mixed-function oxidation assay. PvTPx-1 has two conserved cysteine residues in the amino acid sequence at the positions 50 and 170 which formed a dimer under a non-reducing condition. Using a thiol mixed-function oxidation assay, the antioxidant activity of rPvTPx-1 was revealed. Indirect immunofluorescence microscopy with the specific antibody indicated that PvTPx-1 was expressed in the cytoplasm of the erythrocytic stage of the parasite in a dots-like pattern. The results suggest that P. vivax uses TPx-1 to reduce and detoxify hydrogen peroxides in order to maintain their redox homeostasis and proliferation in the host body.
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
authors.forename: Hassan
authors.initials: H
authors.lastname: Hakimi
authors.forename: Masahito
authors.initials: M
authors.lastname: Asada
authors.forename: Jose Ma M
authors.initials: JM
authors.lastname: Angeles
authors.forename: Noboru
authors.initials: N
authors.lastname: Inoue
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Kawazu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants; D003001:Cloning, Molecular; D019084:Fluorescent Antibody Technique, Indirect; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D054464:Peroxiredoxins / Q000235:genetics / Q000378:metabolism*; D010966:Plasmodium vivax / Q000201:enzymology*; D016133:Polymerase Chain Reaction
chemical_list: D000975:Antioxidants; D054464:Peroxiredoxins