title: Plasmodium vivax and Plasmodium knowlesi: cloning, expression and functional analysis of 1-Cys peroxiredoxin.
abstract: Malaria parasites like other aerobes need to detoxify the reactive oxygen species (ROS) that are mainly produced from hemoglobin degradation in the food vacuole. Since Plasmodium lacks catalase and genuine glutathione peroxidase, they are highly dependent on peroxiredoxins (Prxs) and superoxide dismutases for ROS detoxification. Prxs are protective antioxidant enzymes that act through reduction of hydrogen peroxides. In recent years, several studies have been done on Prx family of human malaria parasites mainly on Plasmodium falciparum but not much on the other human malaria species. In this study 1-Cys peroxiredoxin (1-Cys-Prx) from Plasmodium vivax and Plasmodium knowlesi were cloned and characterized. The complete genes coding for 1-Cys-Prx of P. vivax (Pv1-Cys-Prx) and P. knowlesi (Pk1-Cys-Prx) were PCR amplified and the recombinant proteins were produced by heterologous over-expression in Escherichia coli. Both recombinant proteins showed antioxidant activity with the mixed function oxidation assay. Using specific polyclonal antibodies, it was indicated that Pv1-Cys-Prx and Pk1-Cys-Prx are expressed in the cytoplasm of the parasite. Altogether, the results suggested that 1-Cys-Prxs protect the parasites from oxidative damages.
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. hassanhakimi@gmail.com
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: Satoru
authors.initials: S
authors.lastname: Kawai
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; D003001:Cloning, Molecular; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008251:Macaca; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D006899:Mixed Function Oxygenases / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D054464:Peroxiredoxins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016790:Plasmodium knowlesi / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D010966:Plasmodium vivax / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D011994:Recombinant Proteins; D006899:Mixed Function Oxygenases; D054464:Peroxiredoxins