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A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella

Pathogenic bacteria proliferating inside mammalian host cells need to rapidly adapt to the intracellular environment. How they achieve this and scavenge essential nutrients from the host has been an open question due to the difficulties in distinguishing between bacterial and host metabolites in sit...

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Autores principales: Mitosch, Karin, Beyß, Martin, Phapale, Prasad, Drotleff, Bernhard, Nöh, Katharina, Alexandrov, Theodore, Patil, Kiran R., Typas, Athanasios
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10468081/
https://www.ncbi.nlm.nih.gov/pubmed/37594988
http://dx.doi.org/10.1371/journal.pbio.3002198
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author Mitosch, Karin
Beyß, Martin
Phapale, Prasad
Drotleff, Bernhard
Nöh, Katharina
Alexandrov, Theodore
Patil, Kiran R.
Typas, Athanasios
author_facet Mitosch, Karin
Beyß, Martin
Phapale, Prasad
Drotleff, Bernhard
Nöh, Katharina
Alexandrov, Theodore
Patil, Kiran R.
Typas, Athanasios
author_sort Mitosch, Karin
collection PubMed
description Pathogenic bacteria proliferating inside mammalian host cells need to rapidly adapt to the intracellular environment. How they achieve this and scavenge essential nutrients from the host has been an open question due to the difficulties in distinguishing between bacterial and host metabolites in situ. Here, we capitalized on the inability of mammalian cells to metabolize mannitol to develop a stable isotopic labeling approach to track Salmonella enterica metabolites during intracellular proliferation in host macrophage and epithelial cells. By measuring label incorporation into Salmonella metabolites with liquid chromatography–mass spectrometry (LC–MS), and combining it with metabolic modeling, we identify relevant carbon sources used by Salmonella, uncover routes of their metabolization, and quantify relative reaction rates in central carbon metabolism. Our results underline the importance of the Entner–Doudoroff pathway (EDP) and the phosphoenolpyruvate carboxylase for intracellularly proliferating Salmonella. More broadly, our metabolic labeling strategy opens novel avenues for understanding the metabolism of pathogens inside host cells.
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spelling pubmed-104680812023-08-31 A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella Mitosch, Karin Beyß, Martin Phapale, Prasad Drotleff, Bernhard Nöh, Katharina Alexandrov, Theodore Patil, Kiran R. Typas, Athanasios PLoS Biol Methods and Resources Pathogenic bacteria proliferating inside mammalian host cells need to rapidly adapt to the intracellular environment. How they achieve this and scavenge essential nutrients from the host has been an open question due to the difficulties in distinguishing between bacterial and host metabolites in situ. Here, we capitalized on the inability of mammalian cells to metabolize mannitol to develop a stable isotopic labeling approach to track Salmonella enterica metabolites during intracellular proliferation in host macrophage and epithelial cells. By measuring label incorporation into Salmonella metabolites with liquid chromatography–mass spectrometry (LC–MS), and combining it with metabolic modeling, we identify relevant carbon sources used by Salmonella, uncover routes of their metabolization, and quantify relative reaction rates in central carbon metabolism. Our results underline the importance of the Entner–Doudoroff pathway (EDP) and the phosphoenolpyruvate carboxylase for intracellularly proliferating Salmonella. More broadly, our metabolic labeling strategy opens novel avenues for understanding the metabolism of pathogens inside host cells. Public Library of Science 2023-08-18 /pmc/articles/PMC10468081/ /pubmed/37594988 http://dx.doi.org/10.1371/journal.pbio.3002198 Text en © 2023 Mitosch et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Methods and Resources
Mitosch, Karin
Beyß, Martin
Phapale, Prasad
Drotleff, Bernhard
Nöh, Katharina
Alexandrov, Theodore
Patil, Kiran R.
Typas, Athanasios
A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella
title A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella
title_full A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella
title_fullStr A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella
title_full_unstemmed A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella
title_short A pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular Salmonella
title_sort pathogen-specific isotope tracing approach reveals metabolic activities and fluxes of intracellular salmonella
topic Methods and Resources
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10468081/
https://www.ncbi.nlm.nih.gov/pubmed/37594988
http://dx.doi.org/10.1371/journal.pbio.3002198
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