title: Molecular cloning and biochemical characterization of iron superoxide dismutase from the rodent malaria parasite Plasmodium vinckei.
abstract: Plasmodium parasite utilizes superoxide dismutase family proteins to limit the toxicity of reactive oxygen species, such as produced through hemoglobin degradation. These proteins play an important role in parasite survival during intra-erythrocytic phase. We have identified, and biochemically characterized a putative iron dependent superoxide dismutase from rodent malaria parasite Plasmodium vinckei (PvSOD1). The recombinant PvSOD1 protein was purified to homogeneity through a combination of affinity and gel filtration chromatography. Crosslinking, Native-PAGE and FPLC gel filtration analyses documented that PvSOD1 exists as a dimer in solution, a common feature shared by other Fe-SODs. PvSOD1 is cytosolic in localization and its expression is comparatively higher during trophozoite as compared to that of ring and schizont stages. Enzymatic activity of recombinant PvSOD1 was validated using conventional zymogram analyses and xanthine-xanthine oxidase system. Under optimal conditions, PvSOD1 was highly active and catalyzed the dismutation of superoxide radicals. Furthermore, PvSOD1 showed activity over a broad range of pH and temperature. Inhibition studies suggested that PvSOD1 was inactivated by hydrogen peroxide, and peroxynitrite, but not by cyanide and azide. Since, PvSOD1 plays a central role in oxidative defense mechanism, therefore, characterization of PvSOD1 will be exploited in the screening of new superoxide dismutase inhibitors for their antimalarial activity.
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh 226031, India.
authors.forename: Kirtika
authors.initials: K
authors.lastname: Prakash
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh 226031, India.
authors.forename: Manish
authors.initials: M
authors.lastname: Goyal
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh 226031, India; Academy of Scientific and Innovative Research, (AcSIR), Anusandhan Bhawan, New Delhi, India.
authors.forename: Awakash
authors.initials: A
authors.lastname: Soni
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh 226031, India; Academy of Scientific and Innovative Research, (AcSIR), Anusandhan Bhawan, New Delhi, India.
authors.forename: Arif Jamal
authors.initials: AJ
authors.lastname: Siddiqui
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh 226031, India; Academy of Scientific and Innovative Research, (AcSIR), Anusandhan Bhawan, New Delhi, India.
authors.forename: Jyoti
authors.initials: J
authors.lastname: Bhardwaj
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, Uttar Pradesh 226031, India; Academy of Scientific and Innovative Research, (AcSIR), Anusandhan Bhawan, New Delhi, India. Electronic address: sk_puri@cdri.res.in.
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Puri
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008288:Malaria / Q000469:parasitology*; D008969:Molecular Sequence Data; D018384:Oxidative Stress; D030421:Peroxynitrous Acid / Q000494:pharmacology; D010961:Plasmodium / Q000201:enzymology* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism; D000072105:Superoxide Dismutase-1
chemical_list: D000913:Antibodies, Protozoan; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D030421:Peroxynitrous Acid; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1
keywords: Free radicals; Hydrogen peroxide; Oxidative stress; Plasmodium vinckei; Superoxide dismutase; Zymography