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Título: Prolyl oligopeptidase from Leishmania infantum : biochemical characterization and involvement in macrophage infection
Autor(es): Silva, Camila Lasse
Azevedo, Clênia Santos
Araújo, Carla Nunes de
Motta, Flávia Nader da Silva
Andrade, Milene Aparecida
Rocha, Amanda Pereira
Sampaio, Iracyara
Charneau, Sébastien
Gèze, Marc
Grellier, Phillippe
Santana, Jaime Martins de
Bastos, Izabela Marques Dourado
Afiliação do autor: University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
UMR 7245 MCAM, Musèum National d’Histoire Naturelle, Centre National de la Recherche Scientifique, Paris, France
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
University of Brasília, Faculty of Ceilandia
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
University of Brasília, Faculty of Ceilandia
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
UMR 7245 MCAM, Musèum National d’Histoire Naturelle, Centre National de la Recherche Scientifique, Paris, France
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
University of Brasília, Department of Cell Biology, Laboratory of Protein Chemistry and Biochemistry
UMR 7245 MCAM, Musèum National d’Histoire Naturelle, Centre National de la Recherche Scientifique, Paris, France
CeMIM, Musèum National d’Histoire Naturelle, Paris, France
UMR 7245 MCAM, Musèum National d’Histoire Naturelle, Centre National de la Recherche Scientifique, Paris, France
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
University of Brasília, Department of Cell Biology, Pathogen-Host Interface Laboratory
Assunto: Leishmania infantum
Prolil oligopetidase
Data de publicação: 2020
Editora: Frontiers
Referência: LASSE, CAMILA et al. Prolyl oligopeptidase from Leishmania infantum: biochemical characterization and involvement in macrophage infection. Frontiers in Microbiology, [S. l.], v. 11, 2020. DOI: https://doi.org/10.3389/fmicb.2020.01060. Disponível em: https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2020.01060/full.
Abstract: Leishmania infantum is a flagellated protozoan and one of the main causative agents of visceral leishmaniasis. This disease usually affects the human reticuloendothelial system, can cause death and available therapies may lead to serious side effects. Since it is a neglected tropical disease, the incentives for the development of new drugs are insufficient. It is important to know Leishmania virulence factors that contribute most to the disease in order to develop drugs. In the present work, we have produced L. infantum prolyl oligopeptidase (rPOPLi) in Escherichia coli, and investigated its biochemical properties as well as the effect of POP inhibitors on its enzymatic activity and on the inhibition of the macrophage infection by L. infantum. The optimal activity occurred at pH 7.5 and 37°C in the presence of DTT, the latter increased rPOPLi catalytic efficiency 5-fold on the substrate N-Suc-Gly-Pro-Leu-Gly-Pro-AMC. The enzyme was inhibited by TPCK, TLCK and by two POP specific inhibitors, Z-Pro-prolinal (ZPP, IC50 4.2 nM) and S17092 (IC50 3.5 nM). Besides being a cytoplasmic enzyme, POPLi is also found in punctuate structures within the parasite cytoplasm or associated with the parasite plasma membrane in amastigotes and promastigotes, respectively. Interestingly, S17092 and ZPP prevented parasite invasion in murine macrophages, supporting the involvement of POPLi in the invasive process of L. infantum. These data suggest POPLi as a virulence factor that offers potential as a target for designing new antileishmanial drugs.
Unidade Acadêmica: Instituto de Ciências Biológicas (IB)
Departamento de Biologia Celular (IB CEL)
Faculdade UnB Ceilândia (FCE)
Colegiado de Bases Biológicas e da Saúde (FCE-BASES)
DOI: https://doi.org/10.3389/fmicb.2020.01060
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