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Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi
Over the past three decades, L-proline has become recognized as an important metabolite for trypanosomatids. It is involved in a number of key processes, including energy metabolism, resistance to oxidative and nutritional stress and osmoregulation. In addition, this amino acid supports critical par...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3718742/ https://www.ncbi.nlm.nih.gov/pubmed/23894476 http://dx.doi.org/10.1371/journal.pone.0069419 |
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author | Paes, Lisvane Silva Suárez Mantilla, Brian Zimbres, Flávia Menezes Pral, Elisabeth Mieko Furusho Diogo de Melo, Patrícia Tahara, Erich B. Kowaltowski, Alicia J. Elias, Maria Carolina Silber, Ariel Mariano |
author_facet | Paes, Lisvane Silva Suárez Mantilla, Brian Zimbres, Flávia Menezes Pral, Elisabeth Mieko Furusho Diogo de Melo, Patrícia Tahara, Erich B. Kowaltowski, Alicia J. Elias, Maria Carolina Silber, Ariel Mariano |
author_sort | Paes, Lisvane Silva |
collection | PubMed |
description | Over the past three decades, L-proline has become recognized as an important metabolite for trypanosomatids. It is involved in a number of key processes, including energy metabolism, resistance to oxidative and nutritional stress and osmoregulation. In addition, this amino acid supports critical parasite life cycle processes by acting as an energy source, thus enabling host-cell invasion by the parasite and subsequent parasite differentiation. In this paper, we demonstrate that L-proline is oxidized to Δ(1)-pyrroline-5-carboxylate (P5C) by the enzyme proline dehydrogenase (TcPRODH, E.C. 1.5.99.8) localized in Trypanosoma cruzi mitochondria. When expressed in its active form in Escherichia coli, TcPRODH exhibits a K(m) of 16.58±1.69 µM and a V(max) of 66±2 nmol/min mg. Furthermore, we demonstrate that TcPRODH is a FAD-dependent dimeric state protein. TcPRODH mRNA and protein expression are strongly upregulated in the intracellular epimastigote, a stage which requires an external supply of proline. In addition, when Saccharomyces cerevisiae null mutants for this gene (PUT1) were complemented with the TcPRODH gene, diminished free intracellular proline levels and an enhanced sensitivity to oxidative stress in comparison to the null mutant were observed, supporting the hypothesis that free proline accumulation constitutes a defense against oxidative imbalance. Finally, we show that proline oxidation increases cytochrome c oxidase activity in mitochondrial vesicles. Overall, these results demonstrate that TcPRODH is involved in proline-dependant cytoprotection during periods of oxidative imbalance and also shed light on the participation of proline in energy metabolism, which drives critical processes of the T. cruzi life cycle. |
format | Online Article Text |
id | pubmed-3718742 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-37187422013-07-26 Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi Paes, Lisvane Silva Suárez Mantilla, Brian Zimbres, Flávia Menezes Pral, Elisabeth Mieko Furusho Diogo de Melo, Patrícia Tahara, Erich B. Kowaltowski, Alicia J. Elias, Maria Carolina Silber, Ariel Mariano PLoS One Research Article Over the past three decades, L-proline has become recognized as an important metabolite for trypanosomatids. It is involved in a number of key processes, including energy metabolism, resistance to oxidative and nutritional stress and osmoregulation. In addition, this amino acid supports critical parasite life cycle processes by acting as an energy source, thus enabling host-cell invasion by the parasite and subsequent parasite differentiation. In this paper, we demonstrate that L-proline is oxidized to Δ(1)-pyrroline-5-carboxylate (P5C) by the enzyme proline dehydrogenase (TcPRODH, E.C. 1.5.99.8) localized in Trypanosoma cruzi mitochondria. When expressed in its active form in Escherichia coli, TcPRODH exhibits a K(m) of 16.58±1.69 µM and a V(max) of 66±2 nmol/min mg. Furthermore, we demonstrate that TcPRODH is a FAD-dependent dimeric state protein. TcPRODH mRNA and protein expression are strongly upregulated in the intracellular epimastigote, a stage which requires an external supply of proline. In addition, when Saccharomyces cerevisiae null mutants for this gene (PUT1) were complemented with the TcPRODH gene, diminished free intracellular proline levels and an enhanced sensitivity to oxidative stress in comparison to the null mutant were observed, supporting the hypothesis that free proline accumulation constitutes a defense against oxidative imbalance. Finally, we show that proline oxidation increases cytochrome c oxidase activity in mitochondrial vesicles. Overall, these results demonstrate that TcPRODH is involved in proline-dependant cytoprotection during periods of oxidative imbalance and also shed light on the participation of proline in energy metabolism, which drives critical processes of the T. cruzi life cycle. Public Library of Science 2013-07-22 /pmc/articles/PMC3718742/ /pubmed/23894476 http://dx.doi.org/10.1371/journal.pone.0069419 Text en © 2013 Paes et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Paes, Lisvane Silva Suárez Mantilla, Brian Zimbres, Flávia Menezes Pral, Elisabeth Mieko Furusho Diogo de Melo, Patrícia Tahara, Erich B. Kowaltowski, Alicia J. Elias, Maria Carolina Silber, Ariel Mariano Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi |
title | Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi
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title_full | Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi
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title_fullStr | Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi
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title_full_unstemmed | Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi
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title_short | Proline Dehydrogenase Regulates Redox State and Respiratory Metabolism in Trypanosoma cruzi
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title_sort | proline dehydrogenase regulates redox state and respiratory metabolism in trypanosoma cruzi |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3718742/ https://www.ncbi.nlm.nih.gov/pubmed/23894476 http://dx.doi.org/10.1371/journal.pone.0069419 |
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