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Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging

This study investigates the temporal and spatial interchange of the aromatic amino acid phenylalanine (Phe) between human retinal pigment epithelial cell line (ARPE-19) and tachyzoites of the apicomplexan protozoan parasite Toxoplasma gondii (T. gondii). Stable isotope labelling by amino acids in ce...

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Autores principales: Naemat, Abida, Elsheikha, Hany M., Boitor, Radu A., Notingher, Ioan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4746650/
https://www.ncbi.nlm.nih.gov/pubmed/26857158
http://dx.doi.org/10.1038/srep20811
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author Naemat, Abida
Elsheikha, Hany M.
Boitor, Radu A.
Notingher, Ioan
author_facet Naemat, Abida
Elsheikha, Hany M.
Boitor, Radu A.
Notingher, Ioan
author_sort Naemat, Abida
collection PubMed
description This study investigates the temporal and spatial interchange of the aromatic amino acid phenylalanine (Phe) between human retinal pigment epithelial cell line (ARPE-19) and tachyzoites of the apicomplexan protozoan parasite Toxoplasma gondii (T. gondii). Stable isotope labelling by amino acids in cell culture (SILAC) is combined with Raman micro-spectroscopy to selectively monitor the incorporation of deuterium-labelled Phe into proteins in individual live tachyzoites. Our results show a very rapid uptake of l-Phe(D8) by the intracellular growing parasite. T. gondii tachyzoites are capable of extracting l-Phe(D8) from host cells as soon as it invades the cell. l-Phe(D8) from the host cell completely replaces the l-Phe within T. gondii tachyzoites 7–9 hours after infection. A quantitative model based on Raman spectra allowed an estimation of the exchange rate of Phe as 0.5–1.6 × 10(4) molecules/s. On the other hand, extracellular tachyzoites were not able to consume l-Phe(D8) after 24 hours of infection. These findings further our understanding of the amino acid trafficking between host cells and this strictly intracellular parasite. In particular, this study highlights new aspects of the metabolism of amino acid Phe operative during the interaction between T. gondii and its host cell.
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spelling pubmed-47466502016-02-17 Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging Naemat, Abida Elsheikha, Hany M. Boitor, Radu A. Notingher, Ioan Sci Rep Article This study investigates the temporal and spatial interchange of the aromatic amino acid phenylalanine (Phe) between human retinal pigment epithelial cell line (ARPE-19) and tachyzoites of the apicomplexan protozoan parasite Toxoplasma gondii (T. gondii). Stable isotope labelling by amino acids in cell culture (SILAC) is combined with Raman micro-spectroscopy to selectively monitor the incorporation of deuterium-labelled Phe into proteins in individual live tachyzoites. Our results show a very rapid uptake of l-Phe(D8) by the intracellular growing parasite. T. gondii tachyzoites are capable of extracting l-Phe(D8) from host cells as soon as it invades the cell. l-Phe(D8) from the host cell completely replaces the l-Phe within T. gondii tachyzoites 7–9 hours after infection. A quantitative model based on Raman spectra allowed an estimation of the exchange rate of Phe as 0.5–1.6 × 10(4) molecules/s. On the other hand, extracellular tachyzoites were not able to consume l-Phe(D8) after 24 hours of infection. These findings further our understanding of the amino acid trafficking between host cells and this strictly intracellular parasite. In particular, this study highlights new aspects of the metabolism of amino acid Phe operative during the interaction between T. gondii and its host cell. Nature Publishing Group 2016-02-09 /pmc/articles/PMC4746650/ /pubmed/26857158 http://dx.doi.org/10.1038/srep20811 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Naemat, Abida
Elsheikha, Hany M.
Boitor, Radu A.
Notingher, Ioan
Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging
title Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging
title_full Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging
title_fullStr Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging
title_full_unstemmed Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging
title_short Tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved Raman spectral imaging
title_sort tracing amino acid exchange during host-pathogen interaction by combined stable-isotope time-resolved raman spectral imaging
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4746650/
https://www.ncbi.nlm.nih.gov/pubmed/26857158
http://dx.doi.org/10.1038/srep20811
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