title: Expression of active trypsin-like serine peptidases in the midgut of sugar-feeding female Anopheles aquasalis.
abstract: BACKGROUND
  Anopheles aquasalis is a dipteran of the family Culicidae that is widely distributed in the coastal regions of South and Central America. This species acts as a vector of Plasmodium vivax, an important etiological agent of malaria, which represents a serious public health problem. In mosquitoes, trypsin-like serine proteases are important in blood meal digestion, immune responses and reproductive functions. The study of peptidases expressed in the mosquito midgut is essential to understanding the mechanisms of parasite-host interaction and the physiological process of nutrient digestion.
  
  
  METHODS
  Our study aimed to identify and characterize the proteolytic activities in the midgut of sugar-fed An. aquasalis females using zymographic analyses (substrate-SDS-PAGE), in-solution assays and mass spectrometry.
  
  
  RESULTS
  Here, we used a zymographic analysis to further biochemically characterize the proteolytic profile of the midgut of sugar-feeding An. aquasalis females. The trypsin peptidases migrated between ~17 and ~76 kDa and displayed higher proteolytic activities between pH 7.5 and 10 and at temperatures between 37 °C and 50 °C. Four putative trypsin-like serine peptidases were identified using mass spectrometry and data mining. The molecular masses of these peptidases were similar to those observed using zymography, which suggested that these peptidases could be responsible for some of the observed proteolytic bands.
  
  
  CONCLUSIONS
  Taken together, our results contribute to the gene annotation of the unknown genome of this species, to the tissue location of these peptidases, and to the functional prediction of these crucial enzymes, which all impact further studies of this species.
authors.affiliation: Laboratório de Biologia Molecular e Doenças Endêmicas - Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil. geovane.dl@gmail.com.
authors.forename: Geovane
authors.initials: G
authors.lastname: Dias-Lopes
authors.affiliation: Laboratório de Biologia Molecular e Doenças Endêmicas - Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil. abv@ioc.fiocruz.br.
authors.forename: Andre
authors.initials: A
authors.lastname: Borges-Veloso
authors.affiliation: Laboratório de Pesquisa em Leishmaniose - Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil 4365, Manguinhos, Pav. 26, Sala 509, Rio de Janeiro, Brazil. leovahia@ioc.fiocruz.br.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Saboia-Vahia
authors.affiliation: Laboratório de Química de Proteínas, Departamento de Química, UFRJ, Rio de Janeiro, Brazil. gilberto@iq.ufrj.br.
authors.forename: Gilberto B
authors.initials: GB
authors.lastname: Domont
authors.affiliation: Laboratório de Biologia Molecular e Doenças Endêmicas - Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil. cbritto@ioc.fiocruz.br.
authors.forename: Constança
authors.initials: C
authors.lastname: Britto
authors.affiliation: Laboratório de Pesquisa em Leishmaniose - Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil 4365, Manguinhos, Pav. 26, Sala 509, Rio de Janeiro, Brazil. pcuervo@fiocruz.br.
authors.forename: Patricia
authors.initials: P
authors.lastname: Cuervo
authors.affiliation: Laboratório de Biologia Molecular e Doenças Endêmicas - Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil. jbj@ioc.fiocruz.br.|Departamento de Medicina, Faculdade de Medicina - Universidade Federal de São João Del Rey, São João del Rei, MG, Brazil. jbj@ioc.fiocruz.br.
authors.forename: Jose Batista
authors.initials: JB
authors.lastname: De Jesus
mesh_terms: D000818:Animals; D000852:Anopheles / Q000201:enzymology* / Q000502:physiology; D050260:Carbohydrate Metabolism; D004064:Digestive System / Q000201:enzymology; D005260:Female; D006790:Host-Parasite Interactions; D008288:Malaria / Q000635:transmission*; D010966:Plasmodium vivax / Q000502:physiology*; D059748:Proteolysis; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
chemical_list: C114880:trypsin-like serine protease; D012697:Serine Endopeptidases