Leveraging a Fluorescent Fatty Acid Probe to Discover Cell-Permeable Inhibitors of Plasmodium falciparum Glycerolipid Biosynthesis

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dc.contributor.authorDapper, Christieen
dc.contributor.authorLiu, Jiapengen
dc.contributor.authorKlemba, Michaelen
dc.date.accessioned2023-04-26T17:46:08Zen
dc.date.available2023-04-26T17:46:08Zen
dc.date.issued2022-10en
dc.description.abstractThe human malaria parasite Plasmodium falciparum relies on fatty acid scavenging to supply this essential precursor of lipid synthesis during its asexual replication cycle in human erythrocytes. This dependence on host fatty acids represents a potential vulnerability that can be exploited to develop new anti-malarial therapies. A sensitive and quantitative fluorescence-based approach is presented for characterizing fatty acid acquisition and lipid biosynthesis by asexually replicating, intraerythrocytic Plasmodium falciparum. We show that a BODIPY-containing, green-fluorescent fatty acid analog is efficiently and rapidly incorporated into parasite neutral lipids and phospholipids. Prelabeling with a red-fluorescent ceramide analog permits normalization and enables reliable quantitation of glycerolipid labeling. Inhibition of lipid labeling by competition with natural fatty acids and by acyl-coenzyme A synthetase and diacylglycerol acyltransferase inhibitors demonstrates that the fluorescent fatty acid probe is acquired, activated, and transferred to lipids through physiologically-relevant pathways. To assess its utility in discovering small molecules that block parasite lipid biosynthesis, the lipid labeling assay was used to screen a panel of mammalian lipase inhibitors and a selection of compounds from the "Malaria Box" anti-malarial collection. Several compounds were identified that inhibited the incorporation of the fluorescent fatty acid probe into lipids in cultured parasites at low micromolar concentrations. Two contrasting profiles of suppression of neutral lipid and phospholipid synthesis were observed, which implies the inhibition of distinct pathways. IMPORTANCE The human malaria parasite Plasmodium falciparum relies on fatty acid scavenging to supply this essential precursor of lipid synthesis during its asexual replication cycle in human erythrocytes. This dependence on host fatty acids represents a potential vulnerability that can be exploited to develop new anti-malarial therapies. The quantitative experimental approach described here provides a platform for simultaneously interrogating multiple facets of lipid metabolism- fatty acid uptake, fatty acyl-CoA synthesis, and neutral lipid and phospholipid biosynthesis- and of identifying cell-permeable inhibitors that are active in situ.en
dc.description.notesM. Klemba was supported by HATCH project VA-160082 from the USDA National Institute of Food and Agriculture and by the Biochemistry Department of Virginia Tech. We are grateful to Tapio Nevalainen, University of Eastern Finland, for the generous gift of the MAGL inhibitors described in Aaltonen et al. (38). We thank the Medicines for Malaria Venture for providing the Malaria Box compound collection.en
dc.description.sponsorshipHATCH project from the USDA National Institute of Food and Agriculture [VA-160082]; Biochemistry Department of Virginia Techen
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1128/spectrum.02456-22en
dc.identifier.pmid36314974en
dc.identifier.urihttp://hdl.handle.net/10919/114805en
dc.language.isoenen
dc.publisherAmerican Society for Microbiologyen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectmalariaen
dc.subjectPlasmodium falciparumen
dc.subjectfatty aciden
dc.subjectneutral lipiden
dc.subjectphospholipiden
dc.subjectfluorescent probeen
dc.subjectMalaria Boxen
dc.titleLeveraging a Fluorescent Fatty Acid Probe to Discover Cell-Permeable Inhibitors of Plasmodium falciparum Glycerolipid Biosynthesisen
dc.title.serialMicrobiology Spectrumen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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