Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression

Detalhes bibliográficos
Autor(a) principal: Souza, Raquel Santos de
Data de Publicação: 2019
Outros Autores: Côrtes, Luzia Monteiro de Castro, Charret, Karen dos Santos, Finkelstein, Léa Cysne, Alves, Carlos Roberto, Silva, Franklin Souza
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/35454
Resumo: Fundação Oswaldo Cruz, Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
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spelling Souza, Raquel Santos deCôrtes, Luzia Monteiro de CastroCharret, Karen dos SantosFinkelstein, Léa CysneAlves, Carlos RobertoSilva, Franklin Souza2019-09-10T15:47:43Z2019-09-10T15:47:43Z2019SOUZA, Raquel Santos de et al. Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression. International Journal of Molecular Sciences, v. 20, n. 1315, p. 1-12, 2019.1661-6596https://www.arca.fiocruz.br/handle/icict/3545410.3390/ijms200613151422-0067engMDPILeishmania (Viannia) braziliensisProteaseSerina proteinasesColuna HiTrap benzamidinaLeishmania (Viannia) braziliensisProteaseSerine proteinasesSubtilisin-likeHiTrap benzamidine columnZ-FR-AMCSuc-AFK-AMCSerine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expressioninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz, Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz, Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz, Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz, Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Centro de Desenvolvimento Tecnológico em Saúde. Rio de Janeiro, RJ, Brasil.Serine proteinases in Leishmania (Viannia) braziliensis promastigotes were assessed in this work. This study included the investigation of the enzymatic activity of subcellular fractions obtained from benzamidine affinity chromatography, reverse transcription polymerase chain reactions, and in silico assays of subcellular localization of subtilisin. Promastigote serine proteinases showed gelatinolytic activity with molecular masses of 43 kDa to 170 kDa in the cytosolic fraction and 67 kDa to 170 kDa in the membranous fraction. Serine proteinase activities were detected using N-benzyloxycarbonyl-l-phenylalanyl-l-arginine 7-amino-4-methylcoumarin (Z-FR-AMC) and N-succinyl-l-alanine-l-phenylalanine-l-lysine 7-amino-4-methylcoumarin (Suc-AFK-AMC) as substrates in the cytosolic fraction (Z-FR-AMC = 392 ± 30 µmol.min-1 mg of protein-1 and Suc-AFK-AMC = 252 ± 20 µmol.min-1 mg of protein-1) and in the membranous fraction (Z-FR-AMC = 53 ± 5 µmol.min-1 mg of protein-1 and Suc-AFK-AMC = 63.6 ± 6.5 µmol.min-1 mg of protein-1). Enzyme specificity was shown by inhibition with aprotinin (19% to 80% inhibition) and phenylmethanesulfonyl fluoride (3% to 69%), depending on the subcellular fraction and substrate. The expression of subtilisin (LbrM.13.0860 and LbrM.28.2570) and tryparedoxin peroxidase (LbrM.15.1080) genes was observed by the detection of RNA transcripts 200 bp, 162 bp, and 166 bp long, respectively. Subsequent in silico assays showed LbrM.13.0860 can be located in the cytosol and LbrM.28.2570 in the membrane of the parasite. Data obtained here show the subcellular distribution and expression of serine proteinases, including the subtilisin-like serine proteinases in L. (V.) braziliensis promastigotes.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/35454/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALRaquelSOuza_LuziaCortes_etal_IOC_2019.pdfRaquelSOuza_LuziaCortes_etal_IOC_2019.pdfapplication/pdf3159744https://www.arca.fiocruz.br/bitstream/icict/35454/2/RaquelSOuza_LuziaCortes_etal_IOC_2019.pdf66d8b1d671d06bcd20df2ff676c76c9fMD52TEXTRaquelSOuza_LuziaCortes_etal_IOC_2019.pdf.txtRaquelSOuza_LuziaCortes_etal_IOC_2019.pdf.txtExtracted texttext/plain67630https://www.arca.fiocruz.br/bitstream/icict/35454/3/RaquelSOuza_LuziaCortes_etal_IOC_2019.pdf.txtfba921e11e202aa80b82bc63d3dc4c61MD53icict/354542023-09-14 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dc.title.pt_BR.fl_str_mv Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
title Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
spellingShingle Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
Souza, Raquel Santos de
Leishmania (Viannia) braziliensis
Protease
Serina proteinases
Coluna HiTrap benzamidina
Leishmania (Viannia) braziliensis
Protease
Serine proteinases
Subtilisin-like
HiTrap benzamidine column
Z-FR-AMC
Suc-AFK-AMC
title_short Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
title_full Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
title_fullStr Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
title_full_unstemmed Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
title_sort Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression
author Souza, Raquel Santos de
author_facet Souza, Raquel Santos de
Côrtes, Luzia Monteiro de Castro
Charret, Karen dos Santos
Finkelstein, Léa Cysne
Alves, Carlos Roberto
Silva, Franklin Souza
author_role author
author2 Côrtes, Luzia Monteiro de Castro
Charret, Karen dos Santos
Finkelstein, Léa Cysne
Alves, Carlos Roberto
Silva, Franklin Souza
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Souza, Raquel Santos de
Côrtes, Luzia Monteiro de Castro
Charret, Karen dos Santos
Finkelstein, Léa Cysne
Alves, Carlos Roberto
Silva, Franklin Souza
dc.subject.other.pt_BR.fl_str_mv Leishmania (Viannia) braziliensis
Protease
Serina proteinases
Coluna HiTrap benzamidina
topic Leishmania (Viannia) braziliensis
Protease
Serina proteinases
Coluna HiTrap benzamidina
Leishmania (Viannia) braziliensis
Protease
Serine proteinases
Subtilisin-like
HiTrap benzamidine column
Z-FR-AMC
Suc-AFK-AMC
dc.subject.en.pt_BR.fl_str_mv Leishmania (Viannia) braziliensis
Protease
Serine proteinases
Subtilisin-like
HiTrap benzamidine column
Z-FR-AMC
Suc-AFK-AMC
description Fundação Oswaldo Cruz, Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-09-10T15:47:43Z
dc.date.available.fl_str_mv 2019-09-10T15:47:43Z
dc.date.issued.fl_str_mv 2019
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 SOUZA, Raquel Santos de et al. Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression. International Journal of Molecular Sciences, v. 20, n. 1315, p. 1-12, 2019.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/35454
dc.identifier.issn.pt_BR.fl_str_mv 1661-6596
dc.identifier.doi.none.fl_str_mv 10.3390/ijms20061315
dc.identifier.eissn.none.fl_str_mv 1422-0067
identifier_str_mv SOUZA, Raquel Santos de et al. Serine Proteinases in Leishmania (Viannia) braziliensis Promastigotes Have Distinct Subcellular Distributions and Expression. International Journal of Molecular Sciences, v. 20, n. 1315, p. 1-12, 2019.
1661-6596
10.3390/ijms20061315
1422-0067
url https://www.arca.fiocruz.br/handle/icict/35454
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 MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/35454/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/35454/2/RaquelSOuza_LuziaCortes_etal_IOC_2019.pdf
https://www.arca.fiocruz.br/bitstream/icict/35454/3/RaquelSOuza_LuziaCortes_etal_IOC_2019.pdf.txt
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
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