The first pacifastin elastase inhibitor characterized from a blood sucking animal

Autor(es)

de Marco, Renato ^[1] ; Lovato, Diogo V. ^[1] ; Torquato, Ricardo J. S. ^[1] ; Clara, Renan O. ^[1] ; Buarque, Diego S. ^[1] ; Tanaka, Aparecida S. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Dept Bioquim, BR-04044020 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Peptides; v. 31, n. 7, p. 1280-1286, JUL 2010.
• Citações Web of Science: 11

Resumo

Pacifastin-like protease inhibitors belong to a recent classified protease inhibitor family and they are the smallest protease inhibitors described in animals. In this work, we purified and characterized, for the first time, two neutrophil elastase inhibitors belonging to the pacifastin family from the blood sucking insect Triatoma infestans eggs. The inhibitors showed the same N-terminal sequences, molecular masses of 4257 and 4024 Da by MALDI-TOF mass spectrometry and dissociation constants (Ki) for neutrophil elastase of 0.52 and 0.29 nM, respectively. Using a fat body cDNA library, we cloned a pacifastin precursor containing two protease inhibitor domains similar to locust pacifastins. The first pacifastin domain translated to T. infestans purified protein, named TIPI1. Recombinant TIPI1 expressed in Pichia pastoris system showed similar inhibitory activities compared to the native inhibitor. Its precursor, called TIPP1, is mainly expressed in fat body, and it is up-regulated after blood feeding. The immune challenges of 1(a) instar T. infestans nymph with bacteria or dsRNA strongly stimulated TiPP1 expression in fat body, suggesting a possible role of TiPP1 in T. infestans immunity. This work is the first to characterize a blood feeding insect pacifastin inhibitor. (C) 2010 Elsevier Inc. All rights reserved. (AU)

• Processo FAPESP: 02/13960-8 - Estudos de estrutura e funcao de anti-hemostaticos presentes em artropodes hematofagos.
• Beneficiário: Aparecida Sadae Tanaka
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 09/50434-1 - Estudos moleculares de subtilisinas hipotéticas de Trypanosoma cruzi
• Beneficiário: Diogo Ventura Lovato
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 05/03514-9 - Estudos da função fisiológica e potencial biotecnológico de inibidores de proteases e anti-hemostáticos presentes em artrópodes hematófagos
• Beneficiário: Aparecida Sadae Tanaka
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 05/03339-2 - Estudos bioquimicos e estruturais de inibidores de serinoproteases purificados de ovos do inseto triatoma infestans.
• Beneficiário: Renato de Marco
• Modalidade de apoio: Bolsas no Brasil - Mestrado