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...

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Autores principales:	Lisvane Silva Paes, Brian Suárez Mantilla, Flávia Menezes Zimbres, Elisabeth Mieko Furusho Pral, Patrícia Diogo de Melo, Erich B Tahara, Alicia J Kowaltowski, Maria Carolina Elias, Ariel Mariano Silber
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e812ee8800044e8a96a51ea762a4fe67
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spelling	oai:doaj.org-article:e812ee8800044e8a96a51ea762a4fe672021-11-18T09:03:38ZProline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.1932-620310.1371/journal.pone.0069419https://doaj.org/article/e812ee8800044e8a96a51ea762a4fe672013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23894476/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Over 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 Km of 16.58±1.69 µM and a Vmax 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.Lisvane Silva PaesBrian Suárez MantillaFlávia Menezes ZimbresElisabeth Mieko Furusho PralPatrícia Diogo de MeloErich B TaharaAlicia J KowaltowskiMaria Carolina EliasAriel Mariano SilberPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 7, p e69419 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lisvane Silva Paes Brian Suárez Mantilla Flávia Menezes Zimbres Elisabeth Mieko Furusho Pral Patrícia Diogo de Melo Erich B Tahara Alicia J Kowaltowski Maria Carolina Elias Ariel Mariano Silber Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.
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 Km of 16.58±1.69 µM and a Vmax 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	article
author	Lisvane Silva Paes Brian Suárez Mantilla Flávia Menezes Zimbres Elisabeth Mieko Furusho Pral Patrícia Diogo de Melo Erich B Tahara Alicia J Kowaltowski Maria Carolina Elias Ariel Mariano Silber
author_facet	Lisvane Silva Paes Brian Suárez Mantilla Flávia Menezes Zimbres Elisabeth Mieko Furusho Pral Patrícia Diogo de Melo Erich B Tahara Alicia J Kowaltowski Maria Carolina Elias Ariel Mariano Silber
author_sort	Lisvane Silva Paes
title	Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.
title_short	Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.
title_full	Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.
title_fullStr	Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.
title_full_unstemmed	Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.
title_sort	proline dehydrogenase regulates redox state and respiratory metabolism in trypanosoma cruzi.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/e812ee8800044e8a96a51ea762a4fe67
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Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi.

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