A mevalonate bypass system facilitates elucidation of plastid biology in malaria parasites
Fig 5
Metabolic labeling of PfMev parasites with [2-13C]-mevalonate.
PfMev parasite cultures were treated with 25μM fosmidomycin (50x IC50) and supplemented with either 50μM unlabeled mevalonate or [2-13C]-mevalonate for 48 hours. Metabolites were extracted and analyzed via targeted liquid-chromatography mass spectrometry (LC-MS/MS) using selected reaction monitoring (SRM). A) Detection of IPP/DMAPP (black) in parasites treated with 50μM unlabeled mevalonate and 25μM fosmidomycin, with attempted detection of mass-shifted IPP/DMAPP (red). B) Detection of mass-shifted IPP/DMAPP (red) in parasites treated with 50μM [2-13C]-mevalonate and 25μM fosmidomycin, with attempted detection of IPP/DMAPP (black). C) Detection of FPP (black) in parasites treated with 50μM mevalonate and 25μM fosmidomycin, with attempted detection of mass-shifted FPP (red). D) Detection of mass-shifted FPP (red) in parasites treated with 50μM [2-13C]-mevalonate and 25μM fosmidomycin, with attempted detection of FPP (black). The red asterisks on the biomolecules shown denote the positions of the labeled carbons. Metabolite peaks are labeled with the liquid chromatography column retention time in minutes.