• JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
  • JoomlaWorks Simple Image Rotator
 
  Bookmark and Share
 
 
Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2013.tde-04062014-124916
Document
Author
Full name
Marcell Crispim
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2013
Supervisor
Committee
Silber, Ariel Mariano (President)
Porto, Carlos Roberto Rebello
Thiemann, Otavio Henrique
Title in Portuguese
Caracterização bioquímica e molecular da glutamina sintetase de Trypanosoma cruzi.
Keywords in Portuguese
Trypanosoma cruzi
Amônia
Glutamatos
Glutamina
Metabolismo
Abstract in Portuguese
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.
Title in English
Biochemical and molecular characterization of glutamine synthetase of Trypanosoma cruzi.
Keywords in English
Trypanosoma cruzi
Ammonia
Glutamates
Glutamine
Metabolism
Abstract in English
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.
 
WARNING - Viewing this document is conditioned on your acceptance of the following terms of use:
This document is only for private use for research and teaching activities. Reproduction for commercial use is forbidden. This rights cover the whole data about this document as well as its contents. Any uses or copies of this document in whole or in part must include the author's name.
Publishing Date
2014-06-05
 
WARNING: Learn what derived works are clicking here.
All rights of the thesis/dissertation are from the authors
CeTI-SC/STI
Digital Library of Theses and Dissertations of USP. Copyright © 2001-2024. All rights reserved.