title: 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.
authors.affiliation: Departamento de Farmacia, Facultad de Ciencias, Universidad Nacional de Colombia, Carrera 30 45-03, Bogotá, DC, Colombia.
authors.forename: Giovanny
authors.initials: G
authors.lastname: Garavito
authors.forename: Marie-Carmen
authors.initials: MC
authors.lastname: Monje
authors.forename: Séverine
authors.initials: S
authors.lastname: Maurel
authors.forename: Alexis
authors.initials: A
authors.lastname: Valentin
authors.forename: Françoise
authors.initials: F
authors.lastname: Nepveu
authors.forename: Eric
authors.initials: E
authors.lastname: Deharo
mesh_terms: D000655:Amodiaquine / Q000494:pharmacology; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D000077549:Artemether; D037621:Artemisinins / Q000494:pharmacology; D053626:Atovaquone / Q000494:pharmacology; D002738:Chloroquine / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005978:Glutathione / Q000187:drug effects / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D007202:Indicators and Reagents / Q000737:chemistry; D015767:Mefloquine / Q000494:pharmacology; D008751:Methylene Blue / Q000494:pharmacology; D021261:Parasitic Sensitivity Tests / Q000379:methods*; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000378:metabolism; D011803:Quinine / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D009764:o-Phthalaldehyde / Q000737:chemistry
chemical_list: D000962:Antimalarials; D037621:Artemisinins; D007202:Indicators and Reagents; D000655:Amodiaquine; D006418:Heme; D009764:o-Phthalaldehyde; D002738:Chloroquine; D011803:Quinine; D000077549:Artemether; D005978:Glutathione; D008751:Methylene Blue; D015767:Mefloquine; D053626:Atovaquone