Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolism
During Mycobacterium tuberculosis (Mtb) infection, the virulence factor PtpA belonging to the protein tyrosine phosphatase family is delivered into the cytosol of the macrophage. PtpA interacts with numerous eukaryotic proteins modulating phagosome maturation, innate immune response, apoptosis, and...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-06-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1095060/full |
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| author | Mariana Margenat Gabriela Betancour Vivian Irving Alicia Costábile Tania García-Cedrés María Magdalena Portela María Magdalena Portela Federico Carrión Fernando E. Herrera Andrea Villarino |
| author_facet | Mariana Margenat Gabriela Betancour Vivian Irving Alicia Costábile Tania García-Cedrés María Magdalena Portela María Magdalena Portela Federico Carrión Fernando E. Herrera Andrea Villarino |
| author_sort | Mariana Margenat |
| collection | DOAJ |
| description | During Mycobacterium tuberculosis (Mtb) infection, the virulence factor PtpA belonging to the protein tyrosine phosphatase family is delivered into the cytosol of the macrophage. PtpA interacts with numerous eukaryotic proteins modulating phagosome maturation, innate immune response, apoptosis, and potentially host-lipid metabolism, as previously reported by our group. In vitro, the human trifunctional protein enzyme (hTFP) is a bona fide PtpA substrate, a key enzyme of mitochondrial β-oxidation of long-chain fatty acids, containing two alpha and two beta subunits arranged in a tetramer structure. Interestingly, it has been described that the alpha subunit of hTFP (ECHA, hTFPα) is no longer detected in mitochondria during macrophage infection with the virulent Mtb H37Rv. To better understand if PtpA could be the bacterial factor responsible for this effect, in the present work, we studied in-depth the PtpA activity and interaction with hTFPα. With this aim, we performed docking and in vitro dephosphorylation assays defining the P-Tyr-271 as the potential target of mycobacterial PtpA, a residue located in the helix-10 of hTFPα, previously described as relevant for its mitochondrial membrane localization and activity. Phylogenetic analysis showed that Tyr-271 is absent in TFPα of bacteria and is present in more complex eukaryotic organisms. These results suggest that this residue is a specific PtpA target, and its phosphorylation state is a way of regulating its subcellular localization. We also showed that phosphorylation of Tyr-271 can be catalyzed by Jak kinase. In addition, we found by molecular dynamics that PtpA and hTFPα form a stable protein complex through the PtpA active site, and we determined the dissociation equilibrium constant. Finally, a detailed study of PtpA interaction with ubiquitin, a reported PtpA activator, showed that additional factors are required to explain a ubiquitin-mediated activation of PtpA. Altogether, our results provide further evidence supporting that PtpA could be the bacterial factor that dephosphorylates hTFPα during infection, potentially affecting its mitochondrial localization or β-oxidation activity. |
| first_indexed | 2024-03-13T03:50:24Z |
| format | Article |
| id | doaj.art-0571fb21ae35490b9f1fb55ee0ea9b99 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-03-13T03:50:24Z |
| publishDate | 2023-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-0571fb21ae35490b9f1fb55ee0ea9b992023-06-22T14:08:21ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882023-06-011310.3389/fcimb.2023.10950601095060Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolismMariana Margenat0Gabriela Betancour1Vivian Irving2Alicia Costábile3Tania García-Cedrés4María Magdalena Portela5María Magdalena Portela6Federico Carrión7Fernando E. Herrera8Andrea Villarino9Instituto de Biología, Sección Bioquímica, Facultad de Ciencias-Universidad de la República, Montevideo, UruguayInstituto de Biología, Sección Bioquímica, Facultad de Ciencias-Universidad de la República, Montevideo, UruguayInstituto de Biología, Sección Bioquímica, Facultad de Ciencias-Universidad de la República, Montevideo, UruguayInstituto de Biología, Sección Bioquímica, Facultad de Ciencias-Universidad de la República, Montevideo, UruguayInstituto de Biología, Sección Bioquímica, Facultad de Ciencias-Universidad de la República, Montevideo, UruguayInstituto de Biología, Facultad de Ciencias-Universidad de la República, Montevideo, UruguayUnidad de Bioquímica y Proteómica Analíticas, Institut Pasteur de Montevideo and Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, UruguayLaboratorio de Inmunovirología, Institut Pasteur de Montevideo, Montevideo, UruguayDepartamento de Física, Facultad de Bioquímica y Ciencias Biológicas-Universidad Nacional del Litoral – CONICET, Santa Fe, ArgentinaInstituto de Biología, Sección Bioquímica, Facultad de Ciencias-Universidad de la República, Montevideo, UruguayDuring Mycobacterium tuberculosis (Mtb) infection, the virulence factor PtpA belonging to the protein tyrosine phosphatase family is delivered into the cytosol of the macrophage. PtpA interacts with numerous eukaryotic proteins modulating phagosome maturation, innate immune response, apoptosis, and potentially host-lipid metabolism, as previously reported by our group. In vitro, the human trifunctional protein enzyme (hTFP) is a bona fide PtpA substrate, a key enzyme of mitochondrial β-oxidation of long-chain fatty acids, containing two alpha and two beta subunits arranged in a tetramer structure. Interestingly, it has been described that the alpha subunit of hTFP (ECHA, hTFPα) is no longer detected in mitochondria during macrophage infection with the virulent Mtb H37Rv. To better understand if PtpA could be the bacterial factor responsible for this effect, in the present work, we studied in-depth the PtpA activity and interaction with hTFPα. With this aim, we performed docking and in vitro dephosphorylation assays defining the P-Tyr-271 as the potential target of mycobacterial PtpA, a residue located in the helix-10 of hTFPα, previously described as relevant for its mitochondrial membrane localization and activity. Phylogenetic analysis showed that Tyr-271 is absent in TFPα of bacteria and is present in more complex eukaryotic organisms. These results suggest that this residue is a specific PtpA target, and its phosphorylation state is a way of regulating its subcellular localization. We also showed that phosphorylation of Tyr-271 can be catalyzed by Jak kinase. In addition, we found by molecular dynamics that PtpA and hTFPα form a stable protein complex through the PtpA active site, and we determined the dissociation equilibrium constant. Finally, a detailed study of PtpA interaction with ubiquitin, a reported PtpA activator, showed that additional factors are required to explain a ubiquitin-mediated activation of PtpA. Altogether, our results provide further evidence supporting that PtpA could be the bacterial factor that dephosphorylates hTFPα during infection, potentially affecting its mitochondrial localization or β-oxidation activity.https://www.frontiersin.org/articles/10.3389/fcimb.2023.1095060/fullMycobacterium tuberculosistyrosine phosphatasePtpAhuman mitochondrial trifunctional proteinTFPECHA |
| spellingShingle | Mariana Margenat Gabriela Betancour Vivian Irving Alicia Costábile Tania García-Cedrés María Magdalena Portela María Magdalena Portela Federico Carrión Fernando E. Herrera Andrea Villarino Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolism Frontiers in Cellular and Infection Microbiology Mycobacterium tuberculosis tyrosine phosphatase PtpA human mitochondrial trifunctional protein TFP ECHA |
| title | Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolism |
| title_full | Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolism |
| title_fullStr | Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolism |
| title_full_unstemmed | Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolism |
| title_short | Characteristics of Mycobacterium tuberculosis PtpA interaction and activity on the alpha subunit of human mitochondrial trifunctional protein, a key enzyme of lipid metabolism |
| title_sort | characteristics of mycobacterium tuberculosis ptpa interaction and activity on the alpha subunit of human mitochondrial trifunctional protein a key enzyme of lipid metabolism |
| topic | Mycobacterium tuberculosis tyrosine phosphatase PtpA human mitochondrial trifunctional protein TFP ECHA |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1095060/full |
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