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A non-radiolabeled heme–GSH interaction test for the screening of antimalarial compounds

Abstract : Intraerythrocytic Plasmodium produces large amounts of toxic heme during the digestion of hemoglobin, a parasite specific pathway. Heme is then partially biocristallized into hemozoin and mostly detoxified by reduced glutathione. We proposed an in vitro micro assay to test the ability of drugs to inhibit heme-glutathione dependent degradation. As glutathione and o-phthalaldehyde form a fluorescent adduct, we followed the extinction of the fluorescent signal when heme was added with or without antimalarial compounds. In this assay, 50 microM of amodiaquine, arthemether, chloroquine, methylene blue, mefloquine and quinine inhibited the interaction between glutathione (50 microM) and heme (50 microM), while atovaquone did not. Consequently, this test could detect drugs that can inhibit heme-GSH degradation in a fast, simple and specific way, making it suitable for high throughput screening of potential antimalarials.
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Giovanny Garavito, Marie-Carmen Monje, Séverine Maurel, Alexis Valentin, Françoise Nepveu, et al.. A non-radiolabeled heme–GSH interaction test for the screening of antimalarial compounds. Experimental Parasitology, Elsevier, 2007, 116 (3), pp.311-313. ⟨10.1016/j.exppara.2007.01.005⟩. ⟨hal-02143188⟩

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