In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus

Detalhes bibliográficos
Autor(a) principal: Veloso, André Borges
Data de Publicação: 2015
Outros Autores: Vahia, Leonardo Sabóia, Lopes, Geovane Dias, Domont, Gilberto B., Britto, Constança, Cuervo, Patricia, Jesus, Jose B. de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/13647
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
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spelling Veloso, André BorgesVahia, Leonardo SabóiaLopes, Geovane DiasDomont, Gilberto B.Britto, ConstançaCuervo, PatriciaJesus, Jose B. de2016-04-07T15:55:39Z2016-04-07T15:55:39Z2015VELOSO, André Borges; et al. In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus. Parasites & Vectors, v.8:373, 16p, 2015.1756-3305https://www.arca.fiocruz.br/handle/icict/1364710.1186/s13071-015-0985-0engBioMed CentralIn-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatusinfo: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 Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Química. Laboratório de Química de Proteínas. Unidade de Proteômica. 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 Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil / Universidade Federal de São João del Rei. Faculdade de Medicina. Departamento de Medicina. São João del Rey, MG, Brasil.Background: Culex quinquefasciatus is a hematophagous insect from the Culicidae family that feeds on the blood of humans, dogs, birds and livestock. This species transmits a wide variety of pathogens between humans and animals. The midgut environment is the first location of pathogen-vector interactions for blood-feeding mosquitoes and the expression of specific peptidases in the early stages of feeding could influence the outcome of the infection. Trypsin-like serine peptidases belong to a multi-gene family that can be expressed in different isoforms under distinct physiological conditions. However, the confident assignment of the trypsin genes that are expressed under each condition is still a challenge due to the large number of trypsin-coding genes in the Culicidae family and most likely because they are low abundance proteins. Methods: We used zymography for the biochemical characterization of the peptidase profile of the midgut from C. quinquefasciatus females fed on sugar. Protein samples were also submitted to SDS-PAGE followed by liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis for peptidase identification. The peptidases sequences were analyzed with bioinformatics tools to assess their distinct features. Results: Zymography revealed that trypsin-like serine peptidases were responsible for the proteolytic activity in the midgut of females fed on sugar diet. After denaturation in SDS-PAGE, eight trypsin-like serine peptidases were identified by LC-MS/MS. These peptidases have structural features typical of invertebrate digestive trypsin peptidases but exhibited singularities at the protein sequence level such as: the presence of different amino acids at the autocatalytic motif and substrate binding regions as well as different number of disulfide bounds. Data mining revealed a group of trypsin-like serine peptidases that are specific to C. quinquefasciatus when compared to the culicids genomes sequenced so far. Conclusion: We demonstrated that proteomics approaches combined with bioinformatics tools and zymographic analysis can lead to the functional annotation of trypsin-like serine peptidases coding genes and aid in the understanding of the complexity of peptidase expression in mosquitoes.Culex quinquefasciatusTrypsin-like serine peptidasesZymographyMass spectrometryEspectrometria de MassasPeptídeoCulexTripsinainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; 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dc.title.pt_BR.fl_str_mv In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
title In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
spellingShingle In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
Veloso, André Borges
Culex quinquefasciatus
Trypsin-like serine peptidases
Zymography
Mass spectrometry
Espectrometria de Massas
Peptídeo
Culex
Tripsina
title_short In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
title_full In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
title_fullStr In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
title_full_unstemmed In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
title_sort In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus
author Veloso, André Borges
author_facet Veloso, André Borges
Vahia, Leonardo Sabóia
Lopes, Geovane Dias
Domont, Gilberto B.
Britto, Constança
Cuervo, Patricia
Jesus, Jose B. de
author_role author
author2 Vahia, Leonardo Sabóia
Lopes, Geovane Dias
Domont, Gilberto B.
Britto, Constança
Cuervo, Patricia
Jesus, Jose B. de
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Veloso, André Borges
Vahia, Leonardo Sabóia
Lopes, Geovane Dias
Domont, Gilberto B.
Britto, Constança
Cuervo, Patricia
Jesus, Jose B. de
dc.subject.en.pt_BR.fl_str_mv Culex quinquefasciatus
Trypsin-like serine peptidases
Zymography
Mass spectrometry
topic Culex quinquefasciatus
Trypsin-like serine peptidases
Zymography
Mass spectrometry
Espectrometria de Massas
Peptídeo
Culex
Tripsina
dc.subject.decs.pt_BR.fl_str_mv Espectrometria de Massas
Peptídeo
Culex
Tripsina
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.
publishDate 2015
dc.date.issued.fl_str_mv 2015
dc.date.accessioned.fl_str_mv 2016-04-07T15:55:39Z
dc.date.available.fl_str_mv 2016-04-07T15:55:39Z
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 VELOSO, André Borges; et al. In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus. Parasites & Vectors, v.8:373, 16p, 2015.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/13647
dc.identifier.issn.none.fl_str_mv 1756-3305
dc.identifier.doi.none.fl_str_mv 10.1186/s13071-015-0985-0
identifier_str_mv VELOSO, André Borges; et al. In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus. Parasites & Vectors, v.8:373, 16p, 2015.
1756-3305
10.1186/s13071-015-0985-0
url https://www.arca.fiocruz.br/handle/icict/13647
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 BioMed Central
publisher.none.fl_str_mv BioMed Central
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/13647/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/13647/2/leonardo_vahia_etal_IOC_2015.pdf
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