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Role of Δ1-Pyrroline-5-Carboxylate Dehydrogenase Supports Mitochondrial Metabolism and Host-Cell Invasion ofTrypanosoma cruzi

Mantilla, Brian S.; Paes, Lisvane S.; Pral, Elizabeth M.F.; Martil, Daiana E.; Thiemann, Otavio H.; Fernández-Silva, Patricio; Bastos, Erick L.; Silber, Ariel M.

Role of Δ1-Pyrroline-5-Carboxylate Dehydrogenase Supports Mitochondrial Metabolism and Host-Cell Invasion ofTrypanosoma cruzi Thumbnail


Authors

Brian S. Mantilla

Lisvane S. Paes

Elizabeth M.F. Pral

Daiana E. Martil

Otavio H. Thiemann

Patricio Fernández-Silva

Erick L. Bastos

Ariel M. Silber



Contributors

Abstract

Proline is crucial for energizing critical events throughout the life cycle of Trypanosoma cruzi, the etiological agent of Chagas disease. The proline breakdown pathway consists of two oxidation steps, both of which producereducing equivalents as follows: the conversion of proline to Δ1-pyrroline-5-carboxylate (P5C), and the subsequent conversion of P5C to glutamate. We have identified and characterized the Δ1-pyrroline-5-carboxylate dehydrogenase from T. cruzi (TcP5CDH) and report here on how this enzyme contributes to a central metabolic pathway in this parasite. Size-exclusionchromatography, two-dimensional gel electrophoresis, and small angle x-ray scattering analysis of TcP5CDH revealed an oligomericstate composed of two subunits of six protomers. TcP5CDH was found to complement a yeast strain deficient in PUT2 activity,confirming the enzyme's functional role; and the biochemical parameters (Km, kcat, and kcat/Km) of the recombinant TcP5CDH were determined, exhibiting values comparable with those from T. cruzi lysates. In addition, TcP5CDH exhibited mitochondrial staining during the main stages of the T. cruzi life cycle. mRNA and enzymatic activity levels indicated the up-regulation (6-fold change) of TcP5CDH during the infectivestages of the parasite. The participation of P5C as an energy source was also demonstrated. Overall, we propose that thisenzymatic step is crucial for the viability of both replicative and infective forms of T. cruzi.

Citation

Mantilla, B. S., Paes, L. S., Pral, E. M., Martil, D. E., Thiemann, O. H., Fernández-Silva, P., …Silber, A. M. (2015). Role of Δ1-Pyrroline-5-Carboxylate Dehydrogenase Supports Mitochondrial Metabolism and Host-Cell Invasion ofTrypanosoma cruzi. Journal of Biological Chemistry, 290(12), 7767-7790. https://doi.org/10.1074/jbc.m114.574525

Journal Article Type Article
Online Publication Date Jan 26, 2015
Publication Date Mar 20, 2015
Deposit Date Sep 11, 2019
Publicly Available Date Mar 29, 2024
Journal Journal of Biological Chemistry
Print ISSN 0021-9258
Electronic ISSN 1083-351X
Publisher American Society for Biochemistry and Molecular Biology
Peer Reviewed Peer Reviewed
Volume 290
Issue 12
Pages 7767-7790
DOI https://doi.org/10.1074/jbc.m114.574525

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Copyright Statement
This research was originally published in The Journal of Biological Chemistry. Mantilla, B.S., Paes, L.S., Pral, E.M.F.m Martil, D.E., Thiemann, O.H., Fernández-Silva, P., Bastos, E.L., and Silber, A.M. Role of Δ1-Pyrroline-5-Carboxylate Dehydrogenase Supports Mitochondrial Metabolism and Host-Cell Invasion ofTrypanosoma cruzi. 2015. 290(12): 7767-7790. © the American Society for Biochemistry and Molecular Biology




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