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Abstract

ATP generation in Leishmania donovani amastigote form

Leishmania is the causative agent of various forms of leishmaniasis, a significant cause of morbidity and mortality. The clinical manifestations of the disease range from selfhealing cutaneous and mucocutaneous skin ulcers to a fatal visceral form named visceral leishmaniasis or kala-azar. The differentiation of Leishmania parasites from the insect stage, the promastigote, towards the pathogenic mammalian stage, the amastigote, is triggered primarily by the rise in ambient temperature encountered during the insect to mammal transmission. The survival of amastigote stage is dependent on that of the host. Regarding energy metabolism, which is an essential factor for the survival, parasites adapt to the environment under low oxygen tension in the host using metabolic systems which are very different from that of the host mammals. The amastigote form of L. donovani is independent on oxidative phosphorylation for ATP production. Indeed, its cell growth was not inhibited by 20-fold excess oligomycin and dicyclohexylcarbodiimide, which are the most specific inhibitors of the mitochondrial FoF1-ATP synthase. In contrast, mitochondrial complex I inhibitor rotenone and complex III inhibitor antimycin A inhibited amastigote cell growth, suggesting the role of complex I and complex III in cell survival. Complex II appeared to have no role in cell survival. To further investigate the site of ATP production, we studied the substrate level phosphorylation, which was involved in the synthesis of ATP. Succinate-pyruvate couple showed the highest substrate level phosphorylation, whereas NADHfumarate and NADH-pyruvate couples failed to produce ATP. In contrast, NADPH-fumarate showed the highest rate of ATP formation in promastigotes. We conclude that substrate level phosphorylation is essential for the growth of L. donovani amastigotes.


Author(s): Anup Kumar Roy, Subhasish Mondal, Tanmoy Bera

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