Author | Veloso, André Borges | |
Author | Vahia, Leonardo Sabóia | |
Author | Lopes, Geovane Dias | |
Author | Domont, Gilberto B. | |
Author | Britto, Constança | |
Author | Cuervo, Patricia | |
Author | Jesus, Jose B. de | |
Access date | 2016-04-07T15:55:39Z | |
Available date | 2016-04-07T15:55:39Z | |
Document date | 2015 | |
Citation | 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. | pt_BR |
ISSN | 1756-3305 | |
URI | https://www.arca.fiocruz.br/handle/icict/13647 | |
Language | eng | pt_BR |
Publisher | BioMed Central | pt_BR |
Rights | open access | pt_BR |
Title | In-depth characterization of trypsin-like serine peptidases in the midgut of the sugar fed Culex quinquefasciatus | pt_BR |
Type | Article | pt_BR |
DOI | 10.1186/s13071-015-0985-0 | |
Abstract | 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. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Universidade Federal do Rio de Janeiro. Instituto de Química. Laboratório de Química de Proteínas. Unidade de Proteômica. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Leishmaniose. Rio de Janeiro, RJ, Brasil. | pt_BR |
Affilliation | 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. | pt_BR |
Subject | Culex quinquefasciatus | pt_BR |
Subject | Trypsin-like serine peptidases | pt_BR |
Subject | Zymography | pt_BR |
Subject | Mass spectrometry | pt_BR |
DeCS | Espectrometria de Massas | pt_BR |
DeCS | Peptídeo | pt_BR |
DeCS | Culex | pt_BR |
DeCS | Tripsina | pt_BR |