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Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii

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...

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Autores principales: Kloehn, Joachim, Lunghi, Matteo, Varesio, Emmanuel, Dubois, David, Soldati-Favre, Dominique
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399914/
https://www.ncbi.nlm.nih.gov/pubmed/34436417
http://dx.doi.org/10.3390/metabo11080476
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author Kloehn, Joachim
Lunghi, Matteo
Varesio, Emmanuel
Dubois, David
Soldati-Favre, Dominique
author_facet Kloehn, Joachim
Lunghi, Matteo
Varesio, Emmanuel
Dubois, David
Soldati-Favre, Dominique
author_sort Kloehn, Joachim
collection PubMed
description 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.
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spelling pubmed-83999142021-08-29 Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii Kloehn, Joachim Lunghi, Matteo Varesio, Emmanuel Dubois, David Soldati-Favre, Dominique Metabolites Article 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. MDPI 2021-07-23 /pmc/articles/PMC8399914/ /pubmed/34436417 http://dx.doi.org/10.3390/metabo11080476 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kloehn, Joachim
Lunghi, Matteo
Varesio, Emmanuel
Dubois, David
Soldati-Favre, Dominique
Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii
title Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii
title_full Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii
title_fullStr Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii
title_full_unstemmed Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii
title_short Untargeted Metabolomics Uncovers the Essential Lysine Transporter in Toxoplasma gondii
title_sort untargeted metabolomics uncovers the essential lysine transporter in toxoplasma gondii
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399914/
https://www.ncbi.nlm.nih.gov/pubmed/34436417
http://dx.doi.org/10.3390/metabo11080476
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