Trypsin-like serine peptidase profiles in the egg, larval, and pupal stages of Aedes albopictus

Particle and Fibre Toxicology

Jose B De Jesus¹  Ana Paula de Barros Silva³  Constança Britto³  Geovane Dias-Lopes³  Patricia Cuervo²  Camila Mesquita-Rodrigues³  André Borges-Veloso³  Leonardo Saboia-Vahia³ 

[1] Departamento de Engenharia de Biossistemas, Universidade Federal de São João Del Rey, Minas Gerais, Brazil;Laboratório de Pesquisa em Leishmaniose, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil;Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil

关键词: Zymography;    Peptidases;    Trypsin-like serine peptidases;    Preimaginal stages;    Culicidae;    Aedes albopictus;   

Others  :  1228053
DOI  :  10.1186/1756-3305-6-50

 received in 2012-10-10, accepted in 2013-02-18,  发布年份 2013

【 摘 要 】

Background

Aedes albopictus, a ubiquitous mosquito, is one of the main vectors of dengue and yellow fever, representing an important threat to public health worldwide. Peptidases play key roles in processes such as digestion, oogenesis, and metamorphosis of insects. However, most of the information on the proteolytic enzymes of mosquitoes is derived from insects in the adult stages and is often directed towards the understanding of blood digestion. The aim of this study was to investigate the expression of active peptidases from the preimaginal stages of Ae. albopictus.

Methods

Ae. albopictus eggs, larvae, and pupae were analyzed using zymography with susbtrate-SDS-PAGE. The pH, temperature and peptidase inhibitor sensitivity was evaluated. In addition, the proteolytic activities of larval instars were assayed using the fluorogenic substrate Z-Phe-Arg-AMC.

Results

The proteolytic profile of the larval stage was composed of 8 bands ranging from 17 to 130 kDa. These enzymes displayed activity in a broad range of pH values, from 5.5 to 10.0. The enzymatic profile of the eggs was similar to that of the larvae, although the proteolytic bands of the eggs showed lower intensities. The pupal stage showed a complex proteolytic pattern, with at least 6 bands with apparent molecular masses ranging from 30 to 150 kDa and optimal activity at pH 7.5. Peptidases from larval instars were active from 10°C to 60°C, with optimal activity at temperatures between 37°C and 50°C. The proteolytic profile of both the larval and pupal stages was inhibited by phenyl-methyl sulfonyl-fluoride (PMSF) and Nα-Tosyl L-lysine chloromethyl ketone hydrochloride (TLCK), indicating that the main peptidases expressed during these developmental stages are trypsin-like serine peptidases.

Conclusion

The preimaginal stages of Ae. albopictus exhibited a complex profile of trypsin-like serine peptidase activities. A comparative analysis of the active peptidase profiles revealed differential expression of trypsin-like isoforms among the preimaginal stages, suggesting that some of these enzymes are stage specific. Additionally, a comparison of the peptidase expression between larvae from eggs collected in the natural environment and larvae obtained from the eggs of female mosquitoes maintained in colonies for a long period of time demonstrated that the proteolytic profile is invariable under such conditions.

【 授权许可】

   
2013 Saboia-Vahia et al; licensee BioMed Central Ltd.

【 图 表 】

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