Analysis of the Physiological and Metabolic State of Leishmania Using Heavy Water Labeling
Joachim Kloehn, Malcolm J McConville, PAM Michels (ed.), ML Ginger (ed.), D Zilberstein (ed.)
TRYPANOSOMATIDS | HUMANA PRESS INC | Published : 2020
This protocol describes the use of heavy water (2H2O) labeling to determine the growth rate and metabolic state of Leishmania parasites in culture and in infected animals. In vitro labeling studies are undertaken by cultivating defined parasite developmental stages in standard medium supplemented with 5% 2H2O, resulting in the incorporation of deuterium (2H) into a range of metabolic precursors used in macromolecule (DNA, RNA, protein, lipid, and glycan) synthesis. The rate of turnover of different parasite macromolecules can subsequently be determined by analysis of deuterium enrichment in the different constituents of these macromolecules by gas chromatography-mass spectrometry (GC-MS). To..View full abstract
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Awarded by Australian National Health and Medical Research Council (NHMRC)
This work was supported by the Australian National Health and Medical Research Council (NHMRC Project Grant APP1100000). M.J.M. is an NHMRC Principal Research Fellow.