title: Leishmania donovani mitochondrial iron superoxide dismutase A is released into the cytosol during miltefosine induced programmed cell death.
abstract: The oxidative phosphorylation process is the main source of endogenous reactive oxygen species (ROS) such as superoxide in mitochondria. In mammals, manganese superoxide dismutase plays an important role in detoxification of superoxide before it interferes with mitochondrial function and causes programmed cell death. Here, we investigated the role of Leishmania donovani mitochondrial iron superoxide dismutase-A (LdFeSODA) in protecting the parasite from oxidative stress and in the control of programmed cell death events. We have shown that overexpression of LdFeSODA protects Leishmania donovani from miltefosine induced cytotoxicity and reduced mitochondrial-derived superoxide generation. Furthermore, parasites overexpressing LdFeSODA showed (i) lower level of phosphatidylserine exposure as measured by flow cytometry and fluorescent microscopy; and (ii) reduced level of TUNEL staining of parasites compared to the control parasites. Finally, prolonged incubation of the parasites with miltefosine induced the release of both cytochrome C and LdFeSODA into the cytosol as demonstrated by Western blotting and fluorescence microscopy indicating programmed cell death. The results indicate that LdFeSODA protects the mitochondria of Leishmania from oxidative stress thereby inhibiting programmed cell death.
authors.affiliation: Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada.
authors.forename: Fitsum
authors.initials: F
authors.lastname: Getachew
authors.forename: Lashitew
authors.initials: L
authors.lastname: Gedamu
mesh_terms: D000981:Antiprotozoal Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D045304:Cytochromes c / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology*; D053938:DNA Fragmentation; D007893:Leishmania donovani / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*
chemical_list: D000981:Antiprotozoal Agents; D024101:Mitochondrial Proteins; D015800:Protozoan Proteins; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D010767:Phosphorylcholine; C039128:miltefosine; D045304:Cytochromes c; D013482:Superoxide Dismutase