title: Carbohydrate metabolism in the Toxoplasma gondii apicoplast: localization of three glycolytic isoenzymes, the single pyruvate dehydrogenase complex, and a plastid phosphate translocator.
abstract: Many apicomplexan parasites, such as Toxoplasma gondii and Plasmodium species, possess a nonphotosynthetic plastid, referred to as the apicoplast, which is essential for the parasites' viability and displays characteristics similar to those of nongreen plastids in plants. In this study, we localized several key enzymes of the carbohydrate metabolism of T. gondii to either the apicoplast or the cytosol by engineering parasites which express epitope-tagged fusion proteins. The cytosol contains a complete set of enzymes for glycolysis, which should enable the parasite to metabolize imported glucose into pyruvate. All the glycolytic enzymes, from phosphofructokinase up to pyruvate kinase, are present in the T. gondii genome, as duplicates and isoforms of triose phosphate isomerase, phosphoglycerate kinase, and pyruvate kinase were found to localize to the apicoplast. The mRNA expression levels of all genes with glycolytic products were compared between tachyzoites and bradyzoites; however, a strict bradyzoite-specific expression pattern was observed only for enolase I. The T. gondii genome encodes a single pyruvate dehydrogenase complex, which was located in the apicoplast and absent in the mitochondrion, as shown by targeting of epitope-tagged fusion proteins and by immunolocalization of the native pyruvate dehydrogenase complex. The exchange of metabolites between the cytosol and the apicoplast is likely to be mediated by a phosphate translocator which was localized to the apicoplast. Based on these localization studies, a model is proposed that explains the supply of the apicoplast with ATP and the reduction power, as well as the exchange of metabolites between the cytosol and the apicoplast.
authors: Institute of Medical Microbiology, University of Göttingen, Kreuzbergring 57, D-37075 Göttingen, Germany.
authors: Tobias
authors: T
authors: Fleige
authors: Karsten
authors: K
authors: Fischer
authors: David J P
authors: DJ
authors: Ferguson
authors: Uwe
authors: U
authors: Gross
authors: Wolfgang
authors: W
authors: Bohne
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D050260:Carbohydrate Metabolism; D006019:Glycolysis / Q000502:physiology*; D007527:Isoenzymes / Q000378:metabolism*; D008969:Molecular Sequence Data; D015388:Organelles / Q000378:metabolism*; D028061:Phosphate Transport Proteins / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018087:Plastids / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011768:Pyruvate Dehydrogenase Complex / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D014122:Toxoplasma / Q000166:cytology / Q000201:enzymology / Q000235:genetics / Q000502:physiology
chemical_list: D007527:Isoenzymes; D028061:Phosphate Transport Proteins; D010940:Plant Proteins; D015800:Protozoan Proteins; D011768:Pyruvate Dehydrogenase Complex; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins