Plasmodium falciparum: functional mitochondrial ADP/ATP transporter in Escherichia coli plasmic membrane as a tool for selective drug screening |
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Authors: | Razakantoanina Valérie Florent Isabelle Jaureguiberry Ginette |
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Institution: | Biologie Fonctionnelle des Protozoaires, USM504-EA3335, Département Régulations, Développement, Diversité Moléculaire, Muséum National d'Histoire Naturelle, 61 rue Buffon, 75005 Paris, France. |
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Abstract: | Plasmodium falciparum mitochondrial ADP/ATP transporter or adenylate translocase (PfAdT) was previously characterised at the molecular level and intracellularly located by immuno-electromicroscopy. Inhibition of this transporter blocks parasite development in erythrocytes. In this study, PfAdT was expressed in C43 (DE3) Escherichia coli strain under isopropyl beta-d-thiogalacto-pyranoside (IPTG) induction to screen inhibitory molecules. PfAdT was integrated directly into the bacterial cytoplasmic membrane. Whereas IPTG-induced bacterial cells imported radioactively labelled ATP, non-induced cells did not. The transporter bound specifically ADP and ATP, but not AMP. IPTG-induced cells preloaded with labelled ATP exported ATP after exogenous addition of unlabelled ADP or ATP, indicating a counter exchange transport mechanism. Bongrekic acid and atractyloside, two well-known specific inhibitors of mitochondrial ADP/ATP transporter, were tested. This experimental model was evaluated using three Malagasy crude plants extracts which have shown antiplasmodial activity on in vitro parasite cultures. |
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Keywords: | Malaria Plasmodium falciparum Rickettsia prowazekii Arabidopsis thaliana Brachylaena ramiflora Phyllarthron bernierianum Strychnos sp C43 (DE3) E coli Natural plant extracts ADP/ATP carrier Bongkrekic acid Plants extracts ADP adenosine diphosphate AMP adenosine monophosphate ATP adenosine triphosphate ATR atractyloside BA bongkrekic acid EDTA ethylene diamine tetraacetate IPTG d-thiogalacto-pyranoside" target="_blank">isopropyl beta-d-thiogalacto-pyranoside PfAdT P falciparum adenylate translocase or ADP/ATP transporter PB phosphate buffer SDS sodium dodecyl sulphate YT yeast tryptone |
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