NDLI: Development of a functional assay to detect inhibitors of Plasmodium falciparum glutathione reductase utilizing liquid chromatography-mass spectrometry.

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Development of a functional assay to detect inhibitors of Plasmodium falciparum glutathione reductase utilizing liquid chromatography-mass spectrometry.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Burkard, Lexi Scheuermann, Alexis Simithy, Johayra Calderón, Angela I.
Description	Country affiliation: United States Author Affiliation: Burkard L ( Department of Drug Discovery and Development, Harrison School of Pharmacy, 4306 Walker Building, Auburn University, Auburn, AL, 36849, USA.); Scheuermann A ( Department of Drug Discovery and Development, Harrison School of Pharmacy, 4306 Walker Building, Auburn University, Auburn, AL, 36849, USA.); Simithy J ( Department of Drug Discovery and Development, Harrison School of Pharmacy, 4306 Walker Building, Auburn University, Auburn, AL, 36849, USA.); Calderón AI ( Department of Drug Discovery and Development, Harrison School of Pharmacy, 4306 Walker Building, Auburn University, Auburn, AL, 36849, USA.)
Abstract	Plasmodium falciparum (Pf) like most other organisms, has a sophisticated antioxidant system, part of which includes glutathione reductase (GR). GR works by recycling toxic glutathione disulfide to glutathione, thereby reducing reactive oxygen species and making a form of glutathione (GSH) the parasite can use. Inhibition of this enzyme in Pf impedes parasite growth. In addition, it has been confirmed that PfGR is not identical to human GR. Thus, PfGR is an excellent target for antimalarial drug development. A functional assay utilizing liquid chromatography-mass spectrometry was developed to specifically identify and evaluate inhibitors of PfGR. Using recombinant PfGR enzyme and 1,4-naphthoquinone (1) as a reference compound and 4-nitrobenzothiadiazole (2) and methylene blue (3) as additional compounds, we quantified the concentration of GSH produced compared with a control to determine the inhibitory effect of these compounds. Our results coincide with that presented in literature: compounds 1-3 inhibit PfGR with IC50 values of 2.71, 8.38, and 19.23 µm, respectively. Good precision for this assay was exhibited by low values of intraday and interday coefficient of variation (3.1 and 2.4%, respectively). Thus, this assay can be used to screen for other potential inhibitors of PfGR quickly and accurately.
File Format	HTM / HTML
ISSN	02693879
Issue Number	4
Volume Number	30
e-ISSN	10990801
Journal	Biomedical Chromatography
Language	English
Publisher	Wiley
Publisher Date	2016-04-01
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Analytical Chemistry Discipline Molecular Biology Discipline Biochemistry Discipline Pharmacology Chromatography, Liquid Methods Enzyme Assays Enzyme Inhibitors Pharmacology Glutathione Reductase Antagonists & Inhibitors Glutathione Metabolism Naphthoquinones Plasmodium Falciparum Enzymology Antimalarials Chemistry Humans Malaria, Falciparum Drug Therapy Parasitology Mass Spectrometry Methylene Blue Drug Effects Reproducibility Of Results Thiadiazoles Evaluation Studies Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Medicine Analytical Chemistry Drug Discovery Clinical Biochemistry Biochemistry Molecular Biology Pharmacology

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