1H-NMR metabolite profiles of different strains ofPlasmodium falciparum

Abstract

Although efforts to understand the basis for inter-strain phenotypic variation in the most virulent malaria species, Plasmodium falciparum, have benefited from advances in genomic technologies, there have to date been few metabolomic studies of this parasite. Using 1H-NMR spectroscopy, we have compared the metabolite profiles of red blood cells infected with different P. falciparum strains. These included both chloroquine-sensitive and chloroquine-resistant strains, as well as transfectant lines engineered to express different isoforms of the chloroquine-resistance-conferring pfcrt (P. falciparum chloroquine resistance transporter). Our analyses revealed strain-specific differences in a range of metabolites. There was marked variation in the levels of the membrane precursors choline and phosphocholine, with some strains having >30-fold higher choline levels and >5-fold higher phosphocholine levels than others. Chloroquine-resistant strains showed elevated levels of a number of amino acids relative to chloroquine-sensitive strains, including an approximately 2-fold increase in aspartate levels. The elevation in amino acid levels was attributable to mutations in pfcrt. Pfcrt-linked differences in amino acid abundance were confirmed using alternate extraction and detection (HPLC) methods. Mutations acquired to withstand chloroquine exposure therefore give rise to significant biochemical alterations in the parasite.

Whole Blood Metabolomics by 1H NMR Spectroscopy Provides a New Opportunity To Evaluate Coenzymes and Antioxidants

“…10 In addition, the NADP + /NAD + ratio obtained in our study is similar to that reported for human blood RBCs previously. 28 …”

Combining reverse genetics and nuclear magnetic resonance‐based metabolomics unravels trypanosome‐specific metabolic pathways

“…(Larive et al, 2014). However, these new approaches have not been considered so far for studying parasite metabolisms, except for recent investigations on Plasmodium (Sengupta et al, 2013;Teng et al, 2014). The development of such NMR spectrometry approaches for parasites, including trypanosomes, would certainly complete the toolbox for studying metabolic perturbations in the numerous available mutants affecting metabolic pathways.…”

Bioenergetics-based modeling of Plasmodium falciparum metabolism reveals its essential genes, nutritional requirements, and thermodynamic bottlenecks

“…Nine metabolomics data sets were measured with NMR: eight were obtained from different isolates of P . falciparum trophozoite-infected red blood cells [20] and one from isolated trophozoites [19]. The remaining data set was measured with LC-MS in isolated trophozoites [21].…”