Functional and structural characterization of an ecotin-like serine protease inhibitor from Trypanosoma cruzi

Autor(es):Mostrar menos -

Garcia, Felipe Baena ^[1] ; Cabral, Aline Diniz ^[1] ; Fuhlendorf, Max Mario ^[1] ; da Cruz, Geomar Feitosa ^[2] ; dos Santos, Juliete Vitorino ^[1] ; Ferreira, Graziele Cristina ^[1] ; Carneiro de Rezende, Bernard Robin ^[1] ; Santana, Carla Moreira ^[1] ; Puzer, Luciano ^[1] ; Sasaki, Sergio Daishi ^[1] ; Garcia, Wanius ^[2] ; Speranca, Marcia Aparecida ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed ABC, Ctr Ciencias Nat & Humanas, Rua Acturus 03, Sala 226, Lab 105, BR-09606070 Sao Bernardo Do Campo, SP - Brazil

^[2] Univ Fed ABC, Ctr Ciencias Nat & Humanas, BR-09210170 Santo Andre, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: International Journal of Biological Macromolecules; v. 151, p. 459-466, MAY 15 2020.
• Citações Web of Science: 0

Resumo

Ecotin, a seri ne peptidase inhibitor (ISP), discovered in Escherichia coil, inhibit a wide range of nypsin-like serine peptidases, protecting microorganisms from the host's immune response. In eukaryotes, ISPs encoding genes were found only in Trypanosomatidae protozoa, including the genus Trypanosome, which harbors Trypanosome cruzi, the ethiological agent of Chagas' disease. T. cruzi encodes the ISP2 Trypanosomatidae orthologous, which in Leishmania species present inhibitory activity on mammalian proteases from S1A family suggesting its role in vertebrate-host-parasite interactions. In this study, the structural and biochemical characterization of the recombinant T. cruzi ISP2 (rTdSP2), produced in E. coli was purified in soluble form and analyzed by circular dichroism, fluorescence spectroscopy, native electrophoresis, dynamic light scattering, low X-ray scattering and homology modeling. The obtained data revealed that rTdSP2 was biologically active and forms homodimers in solution. Furthermore, inhibitory activity of rTcISP2 against human neutrophil elastase (HNE) is the highest among ISP2 orthologous from bacteria and trypanosomatids. The role of NE to control T. cruzi parasites through modulation of cellular and humoral innate immune responses in vertebrate hosts, make TcISP2 a key molecular component for parasite infection efficiency, providing a useful basis for investigation of host-parasite interactions and the potential of TcISP2 for biotechnological applications. (C) 2020 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 13/26096-4 - Expressão heteróloga da enzima quitinase de Leishmania (L.) infantum chagasi, Leishmania (V.) braziliensis e Leishmania amazonensis: diagnóstico sorológico e estudo molecular comparativo utilizando sistemas de expressão em células de insetos e bactéria
• Beneficiário: Aline Diniz Cabral
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 17/17275-3 - Estudos do modo de ação de duas mono-oxigenases líticas de polissacarídeos de inseto (ordem isoptera): estrutura molecular, química bioinorgânica e aplicações biotecnológicas
• Beneficiário: Wanius José Garcia da Silva
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 16/14514-4 - Caracterização da quitinase das espécies de Leishmania endêmicas da América do Sul: utilização no diagnóstico em humanos, cães e flebotomíneos.
• Beneficiário: Marcia Aparecida Speranca
• Modalidade de apoio: Auxílio à Pesquisa - Regular