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| Author(s): |
Brian Alejandro Suarez Mantilla
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) |
| Defense date: | 2013-07-10 |
| Examining board members: |
Ariel Mariano Silber;
Jose Roberto Meyer Fernandes;
Paulo Lee Ho;
Alicia Juliana Kowaltowski;
Carsten Wrenger
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| Advisor: | Ariel Mariano Silber |
| Abstract | |
Proline is converted into glutamate through two oxidorreduction steps. First, prolina is oxidized into D1-pyrroline-5-carboxylate (P5C), which is further converted in glutamate by TcP5CDH. In this work we demonstrate that TcP5CDH catalyzes irreversibly the oxidation of gGSA rendering glutamate and NADH/NADPH. The TcP5CDH undergoes a hexameric conformation interacting with inner mitochondrial membranes. Electrons from NADH fed electron respiratory chain for ATP synthesis. Moreover, we show that TcP5CDH is up regulated in infective stages (M e T). When an additional copy of TcP5CDH was ectopically introduced into parasites, mutant cells exhibited morphological changes and alterations in TcGP82. O DSF, reported as inhibitor of aldehyde dehydrogenases, interferes with TcP5CDH, as well as exhibited a trypanocidal effect. RNAi silencing assays for TbP5CDH and developmental expression of this enzyme throughout main insect stages within tsetse fly were also addressed. Our data support biochemical relevance of proline-glutamate pathway in T. cruzi, and constitute first evidences regarding occurrence of P5C metabolism over these trypanosomatid species. (AU) | |