title: Expression of trypsin-like serine peptidases in pre-imaginal stages of Aedes aegypti (Diptera: Culicidae).
abstract: This study reports the biochemical characterization and comparative analyses of highly active serine proteases in the larval and pupal developmental stages of Aedes aegypti (Linnaeus) using substrate-SDS-PAGE. Zymographic analysis of larval stadia detected proteolytic activity in 6-8 bands with apparent molecular masses ranging from 20 to 250 kDa, with activity observed from pH 5.5 to 10.0. The pupal stage showed a complex proteolytic activity in at least 11 bands with apparent Mr ranging from 25 to 250 kDa, and pH optimum at 10.0. The proteolytic activities of both larval and pupal stages were strongly inhibited by phenyl-methyl sulfonyl-fluoride and N-α-Tosyl-L-lysine chloromethyl ketone hydrochloride, indicating that the main proteases expressed by these developmental stages are trypsin-like serine proteases. The enzymes were active at temperatures ranging from 4 to 85°C, with optimal activity between 37 and 60°C, and low activity at 85°C. Comparative analysis between the proteolytic enzymes expressed by larvae and pupae showed that substantial changes in the expression of active trypsin-like serine proteases occur during the developmental cycle of A. aegypti.
authors.affiliation: Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Camila
authors.initials: C
authors.lastname: Mesquita-Rodrigues
authors.forename: Leonardo
authors.initials: L
authors.lastname: Saboia-Vahia
authors.forename: Patrícia
authors.initials: P
authors.lastname: Cuervo
authors.forename: Claudia Masini d'Avila
authors.initials: CM
authors.lastname: Levy
authors.forename: Nildimar Alves
authors.initials: NA
authors.lastname: Honorio
authors.forename: Gilberto Barbosa
authors.initials: GB
authors.lastname: Domont
authors.forename: Jose Batista
authors.initials: JB
authors.lastname: de Jesus
mesh_terms: D000330:Aedes / Q000201:enzymology* / Q000378:metabolism; D000818:Animals; D007814:Larva / Q000201:enzymology / Q000378:metabolism; D008970:Molecular Weight; D010436:Pepstatins / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011679:Pupa / Q000201:enzymology / Q000378:metabolism; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology
chemical_list: D010436:Pepstatins; D010618:Phenanthrolines; D011480:Protease Inhibitors; C114880:trypsin-like serine protease; D014107:Tosyllysine Chloromethyl Ketone; D014108:Tosylphenylalanyl Chloromethyl Ketone; D010664:Phenylmethylsulfonyl Fluoride; D012697:Serine Endopeptidases; C031375:pepstatin; C025205:1,10-phenanthroline