Untargeted metabolomics uncovers the essential lysine transporter in toxoplasma gondii

Article Article scientifique Article Untargeted metabolomics uncovers the essential lysine transporter in toxoplasma gondii eng 2021 author Kloehn Joachim 869011 author Lunghi Matteo 868427 author Varesio Emmanuel 30230 author Dubois David 701488 author Soldati-Favre Dominique 0000-0003-4156-2109 135114 476 10.3390/metabo11080476 34436417 2218-1989 https://www.mdpi.com/2218-1989/11/8/476 Apicomplexan parasites are responsible for devastating diseases, including malaria, toxoplasmosis, and cryptosporidiosis. Current treatments are limited by emerging resistance to, as well as the high cost and toxicity of existing drugs. As obligate intracellular parasites, apicomplexans rely on the uptake of many essential metabolites from their host. Toxoplasma gondii, the causative agent of toxoplasmosis, is auxotrophic for several metabolites, including sugars (e.g., myo-inositol), amino acids (e.g., tyrosine), lipidic compounds and lipid precursors (cholesterol, choline), vitamins, cofactors (thiamine) and others. To date, only few apicomplexan metabolite transporters have been characterized and assigned a substrate. Here, we set out to investigate whether untargeted metabolomics can be used to identify the substrate of an uncharacterized transporter. Based on existing genome- and proteome-wide datasets, we have identified an essential plasma membrane transporter of the major facilitator superfamily in T. gondii-previously termed TgApiAT6-1. Using an inducible system based on RNA degradation, TgApiAT6-1 was depleted, and the mutant parasite's metabolome was compared to that of non-depleted parasites. The most significantly reduced metabolite in parasites depleted in TgApiAT6-1 was identified as the amino acid lysine, for which T. gondii is predicted to be auxotrophic. Using stable isotope-labeled amino acids, we confirmed that TgApiAT6-1 is required for efficient lysine uptake. Our findings highlight untargeted metabolomics as a powerful tool to identify the substrate of orphan transporters Apicomplexa Toxoplasma gondii Amino acids Lysine Major facilitator superfamily Metabolomics Obligate intracellular parasite Solute carrier Stable isotope labeling Transporter Untargeted metabolomics Dewey 615 Dewey 616 European Commission Molecular Basis of Toxoplasma gondii Encystation and Persistence 695596 ToxoPersist EU H2020 Swiss National Science Foundation Cyst wall formation: a persistent challenge in Toxoplasmosis CRSII3_160702 CH Programmes Swiss National Science Foundation IZLIZ3_200277 and CRSII3_160702 CH 31 Section des sciences pharmaceutiques 17 54 Département de microbiologie et médecine moléculaire 21B false 443 Biologie d'un parasite intracellulaire obligatoire 773 Metabolites 11 8 d347c072-fd9c-4979-981d-a42ff4740fad Article (Published version) Public metabolites-11-00476.pdf 2598798 application/pdf CC BY openAccess Faculté des sciences / Section des sciences pharmaceutiques Faculté de médecine / Section de médecine fondamentale / Département de microbiologie et médecine moléculaire

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Untargeted metabolomics uncovers the essential lysine transporter in toxoplasma gondii	Metabolites			2021	336	242

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Dubois, David