Stable isotope labelling of Toxoplasma gondii under iron and copper restriction

Toxoplasma uses metal nutrients like iron and copper for vital functions in their energy metabolism. Here, we have used stable isotope labelling to track the incorporation of glucose and glutamine into metabolic pathways, primarily focusing on central carbon metabolism, when these metals are disrupted. We cultured intracellular Toxoplasma for a period of 4 hours in the present of 13C glucose or 13C glutamine before extracting metabolites and performing mass spectrometry analysis. We included several conditions in our experiment that included metal deprivation (using chelation or genetic means) along with parasites recovering from metal deprivation.