Using a Caesalpinia echinata Lam. protease inhibitor as a tool for studying the roles of neutrophil elastase, cathepsin G and proteinase 3 in pulmonary edema

Using a Caesalpinia echinata Lam. protease inhibitor as a tool for studying the roles of neutrophil elastase, cathepsin G and proteinase 3 in pulmonary edema

Author Cruz-Silva, Ilana Autor UNIFESP Google Scholar
Neuhof, Christiane Google Scholar
Gozzo, Andrezza Justino Autor UNIFESP Google Scholar
Nunes, Viviane Abreu Google Scholar
Hirata, Izaura Yoshico Autor UNIFESP Google Scholar
Sampaio, Misako Uemura Google Scholar
Figueiredo-Ribeiro, Rita de Cassia Google Scholar
Neuhof, Heinz Google Scholar
Araujo, Mariana da Silva Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Univ Giessen
Universidade de São Paulo (USP)
Inst Bot
Abstract Acute lung injury (ALI) is characterized by neutrophil infiltration and the release of proteases, mainly elastase (NE), cathepsin G (Cat G) and proteinase 3 (PR3), which can be controlled by specific endogenous inhibitors. However, inhibitors of these proteases have been isolated from different sources, including plants. for this study, CeEI, or Caesalpinia echinata elastase inhibitor, was purified from C. echinata (Brazil-wood) seeds after acetone fractionation, followed by ion exchange and reversed phase chromatographic steps. Characterization with SDS-PAGE, stability assays, amino acid sequencing and alignment with other protein sequences confirmed that CeEI is a member of the soybean Kunitz trypsin inhibitor family. Like other members of this family, CeEl is a 20 kDa monomeric protein; it is stable within a large pH and temperature range, with four cysteine residues forming two disulfide bridges, conserved amino acid residues and leucine-isoleucine residues in the reactive site. CeEI was able to inhibit NE and Cat G at a nanomolar range (with K(i)s of 1.9 and 3.6 nM, respectively) and inhibited PR3 within a micromolar range (K-i 3.7 mu M), leading to hydrolysis of specific synthetic substrates. in a lung edema model, CeEI reduced the lung weight and pulmonary artery pressure until 180 min after the injection of zymosan-activated polymorphonuclear neutrophils. in experiments performed in the presence of a Cat G and PR3, but not an NE inhibitor, lung edema was reduced only until 150 min and pulmonary artery pressure was similar to that of the control. These results confirm that NE action is crucial to edema establishment and progression. Additionally, CeEI appears to be a useful tool for studying the physiology of pulmonary edema and provides a template for molecular engineering and drug design for ALI therapy. (C) 2013 Elsevier B.V. All rights reserved.
Keywords Caesalpinia echinata Lam
Pulmonary edema
Protease inhibitors
Caesalpinia echinata elastase inhibitor
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundo de Auxilio a Pesquisa da Universidade Federal de São Paulo (FAP/UNIFESP), Brazil
Date 2013-12-01
Published in Phytochemistry. Oxford: Pergamon-Elsevier B.V., v. 96, p. 235-243, 2013.
ISSN 0031-9422 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 235-243
Access rights Closed access
Type Article
Web of Science ID WOS:000328869500023

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