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Metabolite profiling of the intraerythrocytic malaria parasite Plasmodium falciparum by 1 H NMR spectroscopy
Author(s) -
Teng Rongwei,
Junankar Pauline R.,
Bubb William A.,
Rae Caroline,
Mercier Pascal,
Kirk Kiaran
Publication year - 2009
Publication title -
nmr in biomedicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.278
H-Index - 114
eISSN - 1099-1492
pISSN - 0952-3480
DOI - 10.1002/nbm.1323
Subject(s) - nuclear magnetic resonance spectroscopy , metabolite , chloroform , perchloric acid , plasmodium falciparum , chemistry , phosphocholine , chromatography , biochemistry , hypoxanthine , biology , stereochemistry , phospholipid , membrane , organic chemistry , phosphatidylcholine , malaria , immunology , enzyme
NMR spectroscopy was used to identify and quantify compounds in extracts prepared from mature trophozoite‐stage Plasmodium falciparum parasites isolated by saponin‐permeabilisation of the host erythrocyte. One‐dimensional 1 H NMR spectroscopy and four two‐dimensional NMR techniques were used to identify more than 50 metabolites. The intracellular concentrations of over 40 metabolites were estimated from the 1 H NMR spectra of extracts prepared by four extraction methods: perchloric acid, methanol/water, methanol/chloroform/water, and methanol alone. The metabolites quantified included: the majority of the biological α ‐amino acids; 4‐aminobutyric acid; mono‐, di‐ and tri‐carboxylic acids; nucleotides; polyamines; myo ‐inositol; and phosphocholine and phosphoethanolamine. The parasites also contained a significant concentration (up to 12 mM) of the exogenous buffering agent, HEPES. Although the metabolite profiles obtained with each extraction method were broadly similar, perchloric acid was found to have significant advantages over the other extraction media. Copyright © 2009 John Wiley & Sons, Ltd.