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

Author(s)

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]}

Affiliation

^[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

• Document type: Journal article
• Source: Biochemical and Biophysical Research Communications; v. 590, p. 139-144, JAN 29 2022.
• Web of Science Citations: 0

Abstract

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)

• FAPESP's process: 12/03657-8 - Inhibitor and proteases of ectoparasites: relationship of structure-function and identification of the role of these molecules in the interaction of diseases vector e their etiological agents
• Grantee: Aparecida Sadae Tanaka
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 19/03779-5 - Use of phage display as a tool in the diagnosis and control of diseases transmitted by hematophagous vectors
• Grantee: Aparecida Sadae Tanaka
• Support Opportunities: Research Projects - Thematic Grants