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New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state

The bacterial protein tyrosine phosphatase PtpA is a key virulence factor released by Mycobacterium tuberculosis in the cytosol of infected macrophages. So far only two unrelated macrophage components (VPS33B, GSK3α) have been identified as PtpA substrates. As tyrosine phosphatases are capable of us...

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Autores principales: Margenat, Mariana, Labandera, Anne-Marie, Gil, Magdalena, Carrion, Federico, Purificação, Marcela, Razzera, Guilherme, Portela, María Magdalena, Obal, Gonzalo, Terenzi, Hernán, Pritsch, Otto, Durán, Rosario, Ferreira, Ana María, Villarino, Andrea
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5390082/
https://www.ncbi.nlm.nih.gov/pubmed/25743628
http://dx.doi.org/10.1038/srep08819
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author Margenat, Mariana
Labandera, Anne-Marie
Gil, Magdalena
Carrion, Federico
Purificação, Marcela
Razzera, Guilherme
Portela, María Magdalena
Obal, Gonzalo
Terenzi, Hernán
Pritsch, Otto
Durán, Rosario
Ferreira, Ana María
Villarino, Andrea
author_facet Margenat, Mariana
Labandera, Anne-Marie
Gil, Magdalena
Carrion, Federico
Purificação, Marcela
Razzera, Guilherme
Portela, María Magdalena
Obal, Gonzalo
Terenzi, Hernán
Pritsch, Otto
Durán, Rosario
Ferreira, Ana María
Villarino, Andrea
author_sort Margenat, Mariana
collection PubMed
description The bacterial protein tyrosine phosphatase PtpA is a key virulence factor released by Mycobacterium tuberculosis in the cytosol of infected macrophages. So far only two unrelated macrophage components (VPS33B, GSK3α) have been identified as PtpA substrates. As tyrosine phosphatases are capable of using multiple substrates, we developed an improved methodology to pull down novel PtpA substrates from an enriched P-Y macrophage extract using the mutant PtpA D126A. This methodology reduced non-specific protein interactions allowing the identification of four novel putative PtpA substrates by MALDI-TOF-MS and nano LC-MS: three mitochondrial proteins - the trifunctional enzyme (TFP), the ATP synthase, and the sulfide quinone oxidoreductase - and the cytosolic 6-phosphofructokinase. All these proteins play a relevant role in cell energy metabolism. Using surface plasmon resonance, PtpA was found to bind immunopurified human TFP through its catalytic site since TFP-PtpA association was inhibited by a specific phosphatase inhibitor. Moreover, PtpA wt was capable of dephosphorylating immunopurified human TFP in vitro supporting that TFP may be a bona fide PtpA susbtrate. Overall, these results suggest a novel scenario where PtpA-mediated dephosphorylation may affect pathways involved in cell energy metabolism, particularly the beta oxidation of fatty acids through modulation of TFP activity and/or cell distribution.
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spelling pubmed-53900822017-04-14 New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state Margenat, Mariana Labandera, Anne-Marie Gil, Magdalena Carrion, Federico Purificação, Marcela Razzera, Guilherme Portela, María Magdalena Obal, Gonzalo Terenzi, Hernán Pritsch, Otto Durán, Rosario Ferreira, Ana María Villarino, Andrea Sci Rep Article The bacterial protein tyrosine phosphatase PtpA is a key virulence factor released by Mycobacterium tuberculosis in the cytosol of infected macrophages. So far only two unrelated macrophage components (VPS33B, GSK3α) have been identified as PtpA substrates. As tyrosine phosphatases are capable of using multiple substrates, we developed an improved methodology to pull down novel PtpA substrates from an enriched P-Y macrophage extract using the mutant PtpA D126A. This methodology reduced non-specific protein interactions allowing the identification of four novel putative PtpA substrates by MALDI-TOF-MS and nano LC-MS: three mitochondrial proteins - the trifunctional enzyme (TFP), the ATP synthase, and the sulfide quinone oxidoreductase - and the cytosolic 6-phosphofructokinase. All these proteins play a relevant role in cell energy metabolism. Using surface plasmon resonance, PtpA was found to bind immunopurified human TFP through its catalytic site since TFP-PtpA association was inhibited by a specific phosphatase inhibitor. Moreover, PtpA wt was capable of dephosphorylating immunopurified human TFP in vitro supporting that TFP may be a bona fide PtpA susbtrate. Overall, these results suggest a novel scenario where PtpA-mediated dephosphorylation may affect pathways involved in cell energy metabolism, particularly the beta oxidation of fatty acids through modulation of TFP activity and/or cell distribution. Nature Publishing Group 2015-03-06 /pmc/articles/PMC5390082/ /pubmed/25743628 http://dx.doi.org/10.1038/srep08819 Text en Copyright © 2015, Macmillan Publishers Limited. All rights reserved 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 in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Margenat, Mariana
Labandera, Anne-Marie
Gil, Magdalena
Carrion, Federico
Purificação, Marcela
Razzera, Guilherme
Portela, María Magdalena
Obal, Gonzalo
Terenzi, Hernán
Pritsch, Otto
Durán, Rosario
Ferreira, Ana María
Villarino, Andrea
New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state
title New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state
title_full New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state
title_fullStr New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state
title_full_unstemmed New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state
title_short New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state
title_sort new potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase ptpa: hints of a bacterial modulation of macrophage bioenergetics state
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5390082/
https://www.ncbi.nlm.nih.gov/pubmed/25743628
http://dx.doi.org/10.1038/srep08819
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