Use of Radioactive Precursors for Biochemical Characterization the Biosynthesis of Isoprenoids in Intraerythrocytic Stages of Plasmodium falciparum

Kimura, Emı́lia A.; Wunderlich, Gerhard; Jordão, Fabiana Morandi; Tonhosolo, Renata; Gabriel, Heloisa B.; Sussmann, Rodrigo Antonio Ceschini; Saito, Alexandre Yukio; Katzin, Alejandro M.

doi:10.5772/20582

Cited by 1 publication

(1 citation statement)

References 82 publications

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“…Our work focused on biochemical experiments in schizont stages because of previous studies showing a higher incorporation rate of [ 3 H] isoprenic moieties at this stage (Kimura et al, 2011). For this, synchronous cultures of P. falciparum at the ring stage in 25 cm 2 flasks were labelled with either 0.75 μCi/ml [ 3 H] FOH or 40 μCi/ml [ 3 H] isoleucine employed as control of protein synthesis (Martin & Kirk, 2007).…”

Section: Metabolic Labelling Of Parasitesmentioning

confidence: 99%

Beyond the MEP Pathway: a novel kinase required for prenol utilization by malaria parasites

Katzin

Crispim

Verdaguer

et al. 2023

Preprint

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A promising treatment for malaria is a combination of fosmidomycin and clindamycin. Both compounds inhibit the methylerythritol 4-phosphate (MEP) pathway, the parasitic source of farnesyl and geranylgeranyl pyrophosphate (FPP and GGPP, respectively). Both FPP and GGPP are crucial for the biosynthesis of several essential metabolites such as ubiquinone and dolichol, as well as for protein prenylation. Dietary prenols, such as farnesol (FOH) and geranylgeraniol (GGOH), can rescue parasites from MEP inhibitors, suggesting the existence of a missing pathway for prenol salvage via phosphorylation, by competition. In this study, we identified a gene in the genome of P. falciparum, encoding a transmembrane prenol kinase (PolK) involved in the salvage of FOH and GGOH. The enzyme was expressed in Saccharomyces cerevisiae, and its FOH/GGOH kinase activities were experimentally validated. Furthermore, conditional gene knockouts were created to investigate the biological importance of the FOH/GGOH salvage pathway. The knockout parasites were viable but more susceptible to fosmidomycin, and their sensitivity to MEP inhibitors could not be rescued by the addition of prenols. Moreover, the knockout parasites lost their ability to use prenols for protein prenylation. These results demonstrate that FOH/GGOH salvage is an additional source of isoprenoids by malaria parasites when de novo biosynthesis is inhibited. This study also identifies a novel kind of enzyme whose inhibition may potentiate the antimalarial efficacy of drugs that affect isoprenoid metabolism.

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Section: Metabolic Labelling Of Parasitesmentioning

confidence: 99%