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Master's Dissertation
DOI
https://doi.org/10.11606/D.42.2011.tde-13022012-095914
Document
Author
Full name
Jean Douglas Ferraz Cordeiro
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Silber, Ariel Mariano (President)
Cotrim, Paulo Cesar
Thiemann, Otavio Henrique
Title in Portuguese
Clonagem dos genes putativos para prolina desidrogenase e D1-pirrolina-5-carboxilato desidrogenase de Leishmania (Leishmania) amazonensis e caracterização funcional de seus produtos.
Keywords in Portuguese
Leishmania
Clonagem
Enzimas
Oxidação
Abstract in Portuguese
Os tripanosomatídeos utilizam aminoácidos, particularmente a prolina, como fontes de carbono e energia. Essas moléculas estão também envolvidas em outros processos fisiológicos como a metaciclogênesis, osmorregulação, ou resistência a diferentes condições de estresse. Duas enzimas, prolina desidrogenase e D1-pirrolina-5-carboxilato desidrogenase catalisam a oxidação de prolina a glutamato. Embora a presença dessa via em algumas espécies do gênero Leishmania seja amparada por dados do projeto genoma, ainda não ha evidências bioquímicas de sua atividade. Nesse trabalho focamos alguns aspectos dessa via em L. (L.) amazonensis. Demonstramos a presença das ORFs de ambos os genes dessa via (que denominamos LaPRODH e LaP5CDH) e dos seus produtos. Constatamos que ambas enzimas são mitocondriais. Também demonstramos que esses genes complementam cepas de Saccharomyces cerevisiae mutantes para Dput1 ou Dput2, ortólogos de LaPRODH e LaP5CDH respectivamente. Em conjunto, os dados obtidos sugerem a presença e funcionalidade da via prolina-glutamato em L. (L.) amazonensis.
Title in English
Cloning of genes for putatives proline dehydrogenase and D1-pyrroline- 5-carboxylate dehydrogenase from Leishmania (Leishmania) amazonensis and functional characterization of their products.
Keywords in English
Leishmania
Cloning
Enzymes
Oxidation
Abstract in English
Trypanosomatids use amino acids, particularly proline, as a carbon and energy sources. In addition, they are involved in other physiological processes such as metacyclogenesis, osmorregulation or resistance to different stress conditions. Two enzymes, proline dehydrogenase and D1-pyrroline-5-carboxylate dehydrogenase catalyze the oxidation from proline to glutamate. Although, the presence of this pathway in some species of the Leishmania is supported by the genome project, no biochemical data on their activities were shown up to now. In this work we focus on some aspects of the proline-glutamate pathway in L. (L.) amazonensis. We demonstrate the presence of ORFs for both genes (which we called LaPRODH and LaP5CDH) and their products. We also showed that both enzymes are mitochondrial. We could also demonstrate that both genes complement Saccharomyces cerevisiae strains mutant for Dput1 or Dput2, orthologs of LaPRODH and LaP5CDH respectively. Taken together, these data show the presence and functionality of the proline-glutamate pathway in L. (L.) amazonensis.
 
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Publishing Date
2012-03-15
 
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