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Protease expression analysis in recently field-isolated strains of Trypanosoma cruzi: a heterogeneous profile of cysteine protease activities between TC I and TC II major phylogenetic groups

Published online by Cambridge University Press:  14 July 2008

P. FAMPA
Affiliation:
Laboratório de Biologia Molecular de Tripanosomatídeos e Flebotomíneos, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil, 4365, 21045-900, Rio de Janeiro, Brazil
C. V. LISBOA
Affiliation:
Laboratório de Biologia de Tripanosomatídeos, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil, 4365, 21045-900, Rio de Janeiro, Brazil
A. M. JANSEN
Affiliation:
Laboratório de Biologia de Tripanosomatídeos, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil, 4365, 21045-900, Rio de Janeiro, Brazil
A. L. S. SANTOS
Affiliation:
Laboratório de Estudos Integrados em Bioquímica Microbiana, Instituto de Microbiologia, UFRJ, Ilha do Fundão, 21941-590, Rio de Janeiro, Brazil
M. I. RAMIREZ*
Affiliation:
Laboratório de Biologia Molecular de Tripanosomatídeos e Flebotomíneos, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil, 4365, 21045-900, Rio de Janeiro, Brazil
*
*Corresponding author: Laboratório de Biologia Molecular de Tripanosomatídeos e Flebotomíneos, Instituto Oswaldo Cruz, Fiocruz. Av. Brasil, 4365, 21045-900, Rio de Janeiro, Brazil. Tel.: +55 21 38658252. Fax: +55 21 25903495. E-mail address: [email protected]

Summary

Protease expression among TCI and TCII field isolates was analysed. Gelatin-containing gels revealed hydrolysis bands with molecular masses ranging from 45 to 66 kDa. The general protease expression profile showed that TCII isolates presented higher heterogeneity compared to TCI. By utilizing protease inhibitors, we showed that all active proteases at acid pH are cysteine-proteases and all proteases active at alkaline pH are metalloproteases. However, the expression of cruzipain, the T. cruzi major cysteine-protease, did not reproduce a heterogeneous TCII cysteine zymogram profile. Dendogram analyses based on presence/absence matrices of proteases and cruzipain bands showed a TCI separation from the TCII group with 50–60% similarity. We suggest that the observed cysteine protease diversification contributes to differential host infection between TCI and II genotypes.

Type
Original Articles
Copyright
Copyright © 2008 Cambridge University Press

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