Molecular, functional and structural properties of the prolyl oligopeptidase of Trypanosoma cruzi (POP Tc80), which is required for parasite entry into mammalian cells

Molecular, functional and structural properties of the prolyl oligopeptidase of Trypanosoma cruzi (POP Tc80), which is required for parasite entry into mammalian cells

Autor Bastos, Izabela MD Google Scholar
Grellier, Philippe Google Scholar
Martins, Natalia F. Google Scholar
Cadavid-Restrepo, Gloria Google Scholar
Souza-Ault, Marian R. de Google Scholar
Augustyns, Koen Google Scholar
Teixeira, Antonio RL Google Scholar
Schrevel, Joseph Google Scholar
Maigret, Bernard Google Scholar
Silveira, José F. da Autor UNIFESP Google Scholar
Santana, Jaime M. Google Scholar
Instituição Universidade de Brasília (UnB)
Museum Natl Hist Nat
Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
Univ Antwerp
Univ Nancy 1
Universidade Federal de São Paulo (UNIFESP)
Resumo We have demonstrated that the 80 kDa POP Tc80 (prolyl oligopeptidase of Trypanosoma cruzi) is involved in the process of cell invasion, since specific inhibitors block parasite entry into non-phagocytic mammalian host cells. in contrast with other POPs, POP Tc80 is capable of hydrolysing large substrates, such as fibronectin and native collagen. in this study, we present the cloning of the POPTc80 gene, whose deduced amino acid sequence shares considerable identity with other members of the POP family, mainly within its C-terminal portion that forms the catalytic domain. Southern-blot analysis indicated that POPTc80 is present as a single copy in the genome of the parasite. These results are consistent with mapping of POPTc80 to a single chromosome. the active recombinant protein (rPOP Tc80) displayed kinetic properties comparable with those of the native enzyme. Novel inhibitors were assayed with rPOP Tc80, and the most efficient ones presented values of inhibition coefficient K(i) <= 1.52 nM. Infective parasites treated with these specific POP Tc80 inhibitors attached to the surface of mammalian host cells, but were incapable of infecting them. Structural modelling of POP Tc80, based on the crystallized porcine POP, suggested that POP Tc80 is composed of an alpha/beta-hydrolase domain containing the catalytic triad Ser(548)-Asp(631)-His(667). and a seven-bladed beta-propeller non-catalytic domain. Docking analysis suggests that triple-helical collagen access to the catalytic site of POP Tc80 occurs in the vicinity of the interface between the two domains.
Palavra-chave mammalian cell
prolyl oligopeptidase
structural modelling
Tc80 proteinase
Trypanosoma cruzi
trypomastigote
Idioma Inglês
Data de publicação 2005-05-15
Publicado em Biochemical Journal. London: Portland Press Ltd, v. 388, p. 29-38, 2005.
ISSN 0264-6021 (Sherpa/Romeo, fator de impacto)
Publicador Portland Press Ltd
Extensão 29-38
Fonte http://dx.doi.org/10.1042/BJ20041049
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000229464800004
Endereço permanente http://repositorio.unifesp.br/handle/11600/28304

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