versatile inhibitor of digestive enzymes in Aedes aegypti larvae selected from a pacifastin (TiPI) phage display librar

Autor(es)

Manzato, Veronica Moraes ^[1] ; Soares Torquato, Ricardo Jose ^[1] ; Alves Lemos, Francisco Jose ^{[2, 3]} ; Nishiduka, Erika ^[1] ; Tashima, Alexandre Keiji ^[1] ; Tanaka, Aparecida Sadae ^{[3, 1]}

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Dept Bioquim, Escola Paulista Med, Rua 3 Maio 100, BR-04044020 Sao Paulo, SP - Brazil

^[2] Univ Estadual Norte Fluminense, Lab Biotecnol, Rio De Janeiro, RJ - Brazil

^[3] Inst Nacl Ciencia & Tecnol Entomol Mol INCT Em, Rio de Janeiro, RJ - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biochemical and Biophysical Research Communications; v. 590, p. 139-144, JAN 29 2022.
• Citações Web of Science: 0

Resumo

In Brazil, the major vector of arboviruses is Aedes aegypti, which can transmit several alpha and flaviviruses. In this work, a pacifastin protease inhibitor library was constructed and used to select mutants for Ae. aegypti larvae digestive enzymes. The library contained a total of 3.25 x 10(5) cfu with random mutations in the reactive site (P2-P2'). The most successfully selected mutant, TiPI6, a versatile inhibitor, was able to inhibit all three Ae. aegypti larvae proteolytic activities, trypsin-like, chymotrypsin-like and elastase-like activities, with IC50 values of 0.212 nM, 0.107 nM and 0.109 nM, respectively. In conclusion, the TiPI mutated phage display library was shown to be a useful tool for the selection of an inhibitor of proteolytic activities combined in a mix. TiPI6 is capable of controlling all three digestive enzyme activities present in the larval midgut extract. To our knowledge, this is the first time that one inhibitor containing a Gln at the P1 position showed inhibitory activity against trypsin, chymotrypsin, and elastase-like activities. TiPI6 can be a candidate for further larvicidal studies. (C) 2021 Published by Elsevier Inc. (AU)

• Processo FAPESP: 12/03657-8 - Inibidores e proteases de ectoparasitas: relação de estrutura-função e identificação do papel dessas moléculas na interação de vetores de doenças e seus agentes etiológicos
• Beneficiário: Aparecida Sadae Tanaka
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 19/03779-5 - Uso de phage display como ferramenta no diagnóstico e controle de doenças transmitidas por vetores hematófagos
• Beneficiário: Aparecida Sadae Tanaka
• Modalidade de apoio: Auxílio à Pesquisa - Temático