Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes

Detalhes bibliográficos
Autor(a) principal: Morgado-Díaz, José Andrés
Data de Publicação: 2005
Outros Autores: Lopez, Raquel Elisa da Silva, Alves, Carlos Roberto, Soares, Maurilio José, Corte-Real, Suzana, De Simone, Salvatore Giovanni
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/36996
Resumo: Instituto Nacional de Câncer. Centro de Pesquisas. Divisão de Biologia Celular. Grupo de Biologia Estrutural. Rio e Janeiro, RJ, Brasil.
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spelling Morgado-Díaz, José AndrésLopez, Raquel Elisa da SilvaAlves, Carlos RobertoSoares, Maurilio JoséCorte-Real, SuzanaDe Simone, Salvatore Giovanni2019-11-12T12:44:11Z2019-11-12T12:44:11Z2005MORGADO-DIAZ, José Andrés et al. Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes. Memórias do Insituto Oswaldo Cruz, Rio de Janeiro, v. 100, n. 4, p. 377-383, July 2005.0074-0276https://www.arca.fiocruz.br/handle/icict/3699610.1590/s0074-027620050004000071678-8060engFundação Oswaldo Cruz. Instituto Oswaldo CruzLeishmaniaProtease serinaLocalização subcelularTripanosomatídeoLeishmaniaSerine proteaseSubcellular localizationTrypanosomatidSubcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleInstituto Nacional de Câncer. Centro de Pesquisas. Divisão de Biologia Celular. Grupo de Biologia Estrutural. Rio e Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Laboratório de Bioquímica de Proteínas e Peptídios. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Laboratório de Bioquímica de Proteínas e Peptídios. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Ultra-estrutura e Biologia Celular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Ultra-estrutura e Biologia Celular. Rio de Janeiro, RJ, Brasil.Universidade Federal Fluminense. Instituto de Biologia. Departamento de Biologia Celular e Molecular. Niterói, RJ, Brasil.Here we report the subcellular localization of an intracellular serine protease of 68 kDa in axenic promastigotes of Leishmania (Leishmania) amazonensis, using subcellular fractionation, enzymatic assays, immunoblotting, and immunocytochemistry. All fractions were evaluated by transmission electron microscopy and the serine protease activity was measured during the cell fractionation procedure using alpha-N-r-tosyl-L-arginine methyl ester (L-TAME) as substrate, phenylmethylsulphone fluoride (PMSF) and L-1-tosylamino-2-phenylethylchloromethylketone (TPCK) as specific inhibitors. The enzymatic activity was detected mainly in a membranous vesicular fraction (6.5-fold enrichment relative to the whole homogenate), but also in a crude plasma membrane fraction (2.0-fold). Analysis by SDS-PAGE gelatin under reducing conditions demonstrated that the major proteolytic activity was found in a 68 kDa protein in all fractions studied. A protein with identical molecular weight was also recognized in immunoblots by a polyclonal antibody against serine protease (anti-SP), with higher immunoreactivity in the vesicular fraction. Electron microscopic immunolocalization using the same polyclonal antibody showed the enzyme present at the cell surface, as well as in cytoplasmic membranous compartments of the parasite. Our findings indicate that the internal location of this serine protease in L. amazonensis is mainly restricted to the membranes of intracellular compartments resembling endocytic/exocytic elements.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/36996/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALMauriliosoares_SuzanaCReal_etal_IOC_2005.pdfMauriliosoares_SuzanaCReal_etal_IOC_2005.pdfapplication/pdf1393495https://www.arca.fiocruz.br/bitstream/icict/36996/2/Mauriliosoares_SuzanaCReal_etal_IOC_2005.pdf8abb4b05a45e3406f8176e51f5299a6bMD52TEXTMauriliosoares_SuzanaCReal_etal_IOC_2005.pdf.txtMauriliosoares_SuzanaCReal_etal_IOC_2005.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
title Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
spellingShingle Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
Morgado-Díaz, José Andrés
Leishmania
Protease serina
Localização subcelular
Tripanosomatídeo
Leishmania
Serine protease
Subcellular localization
Trypanosomatid
title_short Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
title_full Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
title_fullStr Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
title_full_unstemmed Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
title_sort Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes
author Morgado-Díaz, José Andrés
author_facet Morgado-Díaz, José Andrés
Lopez, Raquel Elisa da Silva
Alves, Carlos Roberto
Soares, Maurilio José
Corte-Real, Suzana
De Simone, Salvatore Giovanni
author_role author
author2 Lopez, Raquel Elisa da Silva
Alves, Carlos Roberto
Soares, Maurilio José
Corte-Real, Suzana
De Simone, Salvatore Giovanni
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Morgado-Díaz, José Andrés
Lopez, Raquel Elisa da Silva
Alves, Carlos Roberto
Soares, Maurilio José
Corte-Real, Suzana
De Simone, Salvatore Giovanni
dc.subject.other.pt_BR.fl_str_mv Leishmania
Protease serina
Localização subcelular
Tripanosomatídeo
topic Leishmania
Protease serina
Localização subcelular
Tripanosomatídeo
Leishmania
Serine protease
Subcellular localization
Trypanosomatid
dc.subject.en.pt_BR.fl_str_mv Leishmania
Serine protease
Subcellular localization
Trypanosomatid
description Instituto Nacional de Câncer. Centro de Pesquisas. Divisão de Biologia Celular. Grupo de Biologia Estrutural. Rio e Janeiro, RJ, Brasil.
publishDate 2005
dc.date.issued.fl_str_mv 2005
dc.date.accessioned.fl_str_mv 2019-11-12T12:44:11Z
dc.date.available.fl_str_mv 2019-11-12T12:44:11Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv MORGADO-DIAZ, José Andrés et al. Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes. Memórias do Insituto Oswaldo Cruz, Rio de Janeiro, v. 100, n. 4, p. 377-383, July 2005.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/36996
dc.identifier.issn.pt_BR.fl_str_mv 0074-0276
dc.identifier.doi.none.fl_str_mv 10.1590/s0074-02762005000400007
dc.identifier.eissn.none.fl_str_mv 1678-8060
identifier_str_mv MORGADO-DIAZ, José Andrés et al. Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania) amazonensis promastigotes. Memórias do Insituto Oswaldo Cruz, Rio de Janeiro, v. 100, n. 4, p. 377-383, July 2005.
0074-0276
10.1590/s0074-02762005000400007
1678-8060
url https://www.arca.fiocruz.br/handle/icict/36996
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Fundação Oswaldo Cruz. Instituto Oswaldo Cruz
publisher.none.fl_str_mv Fundação Oswaldo Cruz. Instituto Oswaldo Cruz
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
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