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Mémoire de Maîtrise
DOI
https://doi.org/10.11606/D.42.2013.tde-04062014-124916
Document
Auteur
Nom complet
Marcell Crispim
Unité de l'USP
Domain de Connaissance
Date de Soutenance
Editeur
São Paulo, 2013
Directeur
Jury
Silber, Ariel Mariano (Président)
Porto, Carlos Roberto Rebello
Thiemann, Otavio Henrique
Titre en portugais
Caracterização bioquímica e molecular da glutamina sintetase de Trypanosoma cruzi.
Mots-clés en portugais
Trypanosoma cruzi
Amônia
Glutamatos
Glutamina
Metabolismo
Resumé en portugais
Trypanosoma cruzi, utiliza os aminoácidos como importantes metabólitos para sua manutenção e viabilidade, dentre eles glutamato e glutamina que participam no suprimento energético, metaciclogênese e síntese de pirimidinas. Neste trabalho a glutamina sintetase de T. cruzi (TcGS) foi estudada sob a hipótese de contribuir com a detoxificação de NH4+. Extratos de T. cruzi e a enzima recombinante pura foram utilizados na caracterização cinética e determinação da conformação oligomérica. Na forma amastigotas, observamos que TcGS é mais expressa e ativa. Além disso, a metionina sulfoximina (MS) inibiu TcGS de maneira competitiva e com alta afinidade e reduziu a proliferação de epimastigotas em excesso extracelular de NH4+, relacionando TcGS com a detoxificação dessa molécula. Assim, o tratamento com MS ao longo do ciclo intracelular reduziu a eclosão de formas tripomastigotas de células CHO-K1, ressaltanto a importância da GS no metabolismo de nitrogênio e no ciclo de vida de T. cruzi.
Titre en anglais
Biochemical and molecular characterization of glutamine synthetase of Trypanosoma cruzi.
Mots-clés en anglais
Trypanosoma cruzi
Ammonia
Glutamates
Glutamine
Metabolism
Resumé en anglais
Trypanosoma cruzi uses amino acids as important metabolites for its maintenance and viability, glutamate and glutamine are involved in energy supply, metacyclogenesis and synthesis of pyrimidines. It was described here the glutamine synthetase of T. cruzi (TcGS), and its relation with the ammonium detoxification. Extracts of T. cruzi and pure recombinant enzyme were used to determine the kinetic parameters and the oligomeric conformation of the enzyme. In amastigote forms, we observed that TcGS is more expressed and active. In addition, methionine sulfoximine (MS) inhibited competitively the TcGS, with high affinity and reduced the epimastigotes proliferation exposed to extracellular ammonium excess, connecting TcGS to the ammonium detoxification. Therefore, the treatment with MS along the intracellular cycle reduced the trypomastigote bursting from the CHO-K1 cells, demonstrating the importance of GS on the T. cruzi life cycle establishment and nitrogen metabolism.
 
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Date de Publication
2014-06-05
 
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