Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos
Trypanosomatids parasites are responsible for neglected diseases in humans and with animal importance. Despite numerous studies, Leishmaniasis and Chagas' disease represent an important public health problem, especially in Brazil. Currently, there are no vaccines for the control of human parasi...
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Tripanossomatídeos Tripanossomatídeos Metaloproteases Metaloproteases gp63 gp63 Gelatina Gelatine Albumina Albumin Zimografia Zymography |
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Tripanossomatídeos Tripanossomatídeos Metaloproteases Metaloproteases gp63 gp63 Gelatina Gelatine Albumina Albumin Zimografia Zymography Silva Júnior, Nilton Jose da Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos |
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Trypanosomatids parasites are responsible for neglected diseases in humans and with animal importance. Despite numerous studies, Leishmaniasis and Chagas' disease represent an important public health problem, especially in Brazil. Currently, there are no vaccines for the control of human parasitic diseases of these diseases and there is still a great need for new drugs or new drug formulations, which are safe, more effective, and less toxic. Metalloproteinases are surface molecules, present in trypanosomatids and are fundamental for parasite-host interaction, changing the homeostasis of the extracellular matrix. Therefore, the aim of this study is to characterize the proteolytic activity of metalloproteases of trypanosomatids of medical importance, causing diseases, such as Chagas' disease and leishmaniasis against substrates. From the axenic cultivation, the total extracts of the parasites were chosen, from which the quantification was made, extracts with the best ones were selected. Protein profiles, as well as enzymatic activity and enzymatic activity in vitro of different crude extracts of the Trypanosomatids, were evaluated. In all assays performed by SDS-PAGE, a clear distinction was observed between the parasite profiles. Through zymography tests, the proteolytic profile of the proteins in the crude extract of Leishmania spp. on the gelatin and albumin substrate. Regarding Trypanosoma cruzi, a proteolytic pattern similar to that described in the literature was found for cruzipain. These proteases were subjected to degradation tests against serum albumin substrates, at different time intervals, using the SDS- PAGE method, where it was possible to verify the affinity of the proteolytic activity profile of the species selected for this study. |
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Silva, Marcelo Sousa |
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Silva, Marcelo Sousa Silva Júnior, Nilton Jose da |
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bachelorThesis |
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Silva Júnior, Nilton Jose da |
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Silva Júnior, Nilton Jose da |
title |
Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos |
title_short |
Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos |
title_full |
Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos |
title_fullStr |
Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos |
title_full_unstemmed |
Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos |
title_sort |
estudo de interação bioquímica da metaloproteases de leishmania spp. e trypanosoma cruzi com substratos proteicos |
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Universidade Federal do Rio Grande do Norte |
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2021 |
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https://repositorio.ufrn.br/handle/123456789/35702 |
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AT silvajuniorniltonjoseda estudodeinteracaobioquimicadametaloproteasesdeleishmaniasppetrypanosomacruzicomsubstratosproteicos |
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ri-123456789-357022021-09-28T16:44:53Z Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos Silva Júnior, Nilton Jose da Silva, Marcelo Sousa Cláudia Gonçalves Moreno Donato, Silvia Tavares Oliveira, Johny Wisllas Tripanossomatídeos Tripanossomatídeos Metaloproteases Metaloproteases gp63 gp63 Gelatina Gelatine Albumina Albumin Zimografia Zymography Trypanosomatids parasites are responsible for neglected diseases in humans and with animal importance. Despite numerous studies, Leishmaniasis and Chagas' disease represent an important public health problem, especially in Brazil. Currently, there are no vaccines for the control of human parasitic diseases of these diseases and there is still a great need for new drugs or new drug formulations, which are safe, more effective, and less toxic. Metalloproteinases are surface molecules, present in trypanosomatids and are fundamental for parasite-host interaction, changing the homeostasis of the extracellular matrix. Therefore, the aim of this study is to characterize the proteolytic activity of metalloproteases of trypanosomatids of medical importance, causing diseases, such as Chagas' disease and leishmaniasis against substrates. From the axenic cultivation, the total extracts of the parasites were chosen, from which the quantification was made, extracts with the best ones were selected. Protein profiles, as well as enzymatic activity and enzymatic activity in vitro of different crude extracts of the Trypanosomatids, were evaluated. In all assays performed by SDS-PAGE, a clear distinction was observed between the parasite profiles. Through zymography tests, the proteolytic profile of the proteins in the crude extract of Leishmania spp. on the gelatin and albumin substrate. Regarding Trypanosoma cruzi, a proteolytic pattern similar to that described in the literature was found for cruzipain. These proteases were subjected to degradation tests against serum albumin substrates, at different time intervals, using the SDS- PAGE method, where it was possible to verify the affinity of the proteolytic activity profile of the species selected for this study. Os parasitas do grupo dos Tripanosomatídeos são responsáveis por importantes doenças negligenciadas de importância humana e animal. Apesar de inúmeros estudos, a Leishmaniose e a Doença de Chagas representam um importante problema de saúde pública, principalmente no Brasil. Atualmente, não existe vacinas para o controle das doenças parasitárias humanas e ainda existe uma grande necessidade de novos fármacos ou novas formulações dos fármacos, seguros, com maior eficácia e menos tóxicos. As metaloproteases são moléculas de superfície, presente nos tripanossomatídeos sendo fundamentais para interação parasita-hospedeiro, alterando a homeostasia da matriz extracelular. Diante disso, o objetivo deste estudo é a caracterização da atividade proteolítica de metaloproteases de Tripanosomatídeos de importância médica, causadoras de doenças, tais como a Doença de Chagas e as leishmanioses frente a substratos. A partir do cultivo axênico foram obtidos os extratos totais dos parasitas, do qual foi feito a quantificação, selecionando extratos com as melhores concentrações. Foram avaliados os perfis proteicos, bem como a atividade enzimática e atividade enzimática in vitro de diferentes extratos brutos dos Tripanosomatídeos. Através de ensaios por SDS-PAGE foi observado clara distinção entre os perfis dos parasitas. Através de ensaios de zimografia foi confirmado o perfil proteolítico das proteínas do extrato bruto de Leishmania spp. sobre o substrato gelatina e albumina. Em relação ao Trypanosoma cruzi, um padrão proteolítico semelhante com o descrito na literatura foi encontrado para a cruzipaína. Essas proteases foram submetidas a ensaios de degradação frente a substratos de albumina sérica, em diferentes intervalos de tempo, usando o método de SDS-PAGE, onde foi possível verificar a afinidade o perfil de atividade proteolítica das espécies selecionadas para esse estudo. 2021-04-23T19:28:37Z 2021-09-20T17:49:23Z 2021-04-23T19:28:37Z 2021-09-20T17:49:23Z 2021-03-05 bachelorThesis 20160153472 SILVA JUNIOR, Nilton José da. Estudo de interação bioquímica da metaloproteases de Leishmania spp. e Trypanosoma cruzi com substratos proteicos. 2021. 59 f. Trabalho de Conclusão de Curso (Graduação em Farmácia) - Departamento de Farmácia, Universidade Federal do Rio Grande do Norte, Natal, 2021. https://repositorio.ufrn.br/handle/123456789/35702 pt_BR Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ application/octet-stream Universidade Federal do Rio Grande do Norte Brasil UFRN Farmácia |