The first characterization of a cystatin and a cathepsin L-like peptidase from Aedes aegypti and their possible role in DENV infection by the modulation of apoptosis.

Autor: Oliveira FAA; Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), Brazil., Buri MV; Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), Brazil., Rodriguez BL; Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), Brazil., Costa-da-Silva AL; Department of Parasitology, Instituto de Ciências Biomédicas, Universidade de São Paulo (USP), Brazil., Araújo HRC; Department of Parasitology, Instituto de Ciências Biomédicas, Universidade de São Paulo (USP), Brazil., Capurro ML; Department of Parasitology, Instituto de Ciências Biomédicas, Universidade de São Paulo (USP), Brazil., Lu S; Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), Brazil. Electronic address: lu.stephen@unifesp.br., Tanaka AS; Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), Brazil. Electronic address: astanaka10@unifesp.br.
Jazyk: angličtina
Zdroj: International journal of biological macromolecules [Int J Biol Macromol] 2020 Mar 01; Vol. 146, pp. 141-149. Date of Electronic Publication: 2019 Dec 16.
DOI: 10.1016/j.ijbiomac.2019.12.010
Abstrakt: Recently, a salivary gland transcriptome study demonstrated that the transcripts of a putative cystatin gene (SeqID AAEL013287; Aacystatins) from Aedes aegypti were increased in DENV2-infected mosquitoes and that silencing of the Aacystatin gene resulted in an increase in DENV titres. In this work, Aacystatin was biochemically characterized; the purified recombinant inhibitor was able to inhibit typical cysteine proteases with a Ki in the nM range. Pulldown assays using Aag2 cell extracts identified a cathepsin L-like peptidase (AaCatL) as a possible target of Aacystatin. Purified recombinant AaCatL had an optimal pH of 5.0 and displayed a preference for Leu, Val and Phe residues at P2, which is common for other cathepsin L-like peptidases. Transcription analysis of Aacystatin and AaCatL in the salivary glands and midgut of DENV2-infected mosquitoes revealed a negative correlation between DENV2 titres and levels of the inhibitor and peptidase, suggesting their involvement in DENV2-mosquito interactions. Considering that apoptosis may play an important role during viral infections, the possible involvement of Aacystatin in staurosporine-induced apoptosis in Aag2 cells was investigated; the results showed higher expression of the inhibitor in treated cells; moreover, pre incubation with rAacystatin was able to increase Aag2 cell viability.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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