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A zymographic study of metalloprotease activities in extracts and extracellular secretions of Leishmania (Viannia) braziliensis strains

Published online by Cambridge University Press:  03 October 2005

P. CUERVO
Affiliation:
Departamento de Medicina Tropical, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil Centro Internacional de Entrenamiento e Investigaciones Médicas CIDEIM, Cali, Colombia Departamento de Imunologia, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil
L. SABÓIA-VAHIA
Affiliation:
Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil Instituto de Biofisica, Carlos Chagas Filho, UFRJ, Rio de Janeiro, Brasil
F. COSTA SILVA-FILHO
Affiliation:
Instituto de Biofisica, Carlos Chagas Filho, UFRJ, Rio de Janeiro, Brasil
O. FERNANDES
Affiliation:
Departamento de Medicina Tropical, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil
E. CUPOLILLO
Affiliation:
Departamento de Imunologia, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil
J. B. DE JESUS
Affiliation:
Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil Instituto de Biofisica, Carlos Chagas Filho, UFRJ, Rio de Janeiro, Brasil

Abstract

Proteolytic activities of 5 strains of Leishmania (Viannia) braziliensis isolated from Brazilian and Colombian patients, presenting distinct clinical manifestations, were characterized and compared using whole-promastigote extracts and extracellular secretions. Zymographic assays concerning whole-cell extracts and supernatants resulted in the detection of high molecular weight bands, ranging from 50 to 125 kDa. Proteolytic activities from both whole-cell extracts and supernatants were optimal in a pH range 5·5 to 9·0 for all analysed strains. Such protease activities were inhibited when 10 mM 1,10-phenanthroline was assayed, strongly suggesting that the enzymes responsible for hydrolysis of the substrate belong to the metalloproteases class. Distinct profiles of metalloproteases were observed among the studied L. (V.) braziliensis strains. Differences among the microorganisms might be related to the geographical origin of the strains and/or to the clinical presentation.

Type
Research Article
Copyright
2005 Cambridge University Press

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