Busca avançada
Ano de início
Entree
(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Role of Delta(1)-Pyrroline-5-Carboxylate Dehydrogenase Supports Mitochondrial Metabolism and Host-Cell Invasion of Trypanosoma cruzi

Texto completo
Autor(es):
Mantilla, Brian S. [1] ; Paes, Lisvane S. [1] ; Pral, Elizabeth M. F. [1] ; Martil, Daiana E. [2] ; Thiemann, Otavio H. [2] ; Fernandez-Silva, Patricio [3] ; Bastos, Erick L. [4] ; Silber, Ariel M. [1]
Número total de Autores: 8
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Inst Ciencias Biomed, Dept Parasitol, BR-05508000 Sao Paulo - Brazil
[2] Univ Sao Paulo, Inst Fis Sao Carlos, Lab Biol Estrutural, BR-13560590 Sao Paulo - Brazil
[3] Univ Zaragoza, Dept Bioquim Biol Mol & Celular, E-50013 Zaragoza - Spain
[4] Univ Sao Paulo, Inst Quim, Dept Quim Fundamental, BR-13560590 Sao Paulo - Brazil
Número total de Afiliações: 4
Tipo de documento: Artigo Científico
Fonte: Journal of Biological Chemistry; v. 290, n. 12, p. 7767-7790, MAR 20 2015.
Citações Web of Science: 21
Resumo

Background: The enzyme, (1)-pyrroline-5-carboxylate dehydrogenase (P5CDH), is a key enzyme involved in proline catabolism. Results: The parasite Trypanosoma cruzi up-regulates P5CDH during host infection. Conclusion:T. cruzi uses P5C to produce energy, resist metabolic stress, and invade host cells through this mitochondrion-bound enzyme. Significance: The oxidation of P5C is sufficient to supply energy and enhance the pathogenicity of T. cruzi. 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 produce reducing equivalents as follows: the conversion of proline to (1)-pyrroline-5-carboxylate (P5C), and the subsequent conversion of P5C to glutamate. We have identified and characterized the (1)-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-exclusion chromatography, two-dimensional gel electrophoresis, and small angle x-ray scattering analysis of TcP5CDH revealed an oligomeric state 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 (K-m, k(cat), and k(cat)/K-m) 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 infective stages of the parasite. The participation of P5C as an energy source was also demonstrated. Overall, we propose that this enzymatic step is crucial for the viability of both replicative and infective forms of T. cruzi. (AU)

Processo FAPESP: 13/18970-6 - Caracterização do efeito Disulfiram em Trypanosoma cruzi
Beneficiário:Ariel Mariano Silber
Linha de fomento: Auxílio à Pesquisa - Regular