Parasitology



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


P. CUERVO a1a2a5, L. SABÓIA-VAHIA a3a4, F. COSTA SILVA-FILHO a4, O. FERNANDES a1, E. CUPOLILLO a5 and J. B. DE JESUS a3a4c1
a1 Departamento de Medicina Tropical, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil
a2 Centro Internacional de Entrenamiento e Investigaciones Médicas CIDEIM, Cali, Colombia
a3 Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil
a4 Instituto de Biofisica, Carlos Chagas Filho, UFRJ, Rio de Janeiro, Brasil
a5 Departamento de Imunologia, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brasil

Article author query
cuervo p   [PubMed][Google Scholar] 
saboia-vahia l   [PubMed][Google Scholar] 
costa silva-filho f   [PubMed][Google Scholar] 
fernandes o   [PubMed][Google Scholar] 
cupolillo e   [PubMed][Google Scholar] 
de jesus jb   [PubMed][Google Scholar] 

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.

(Received April 13 2005)
(Revised June 24 2005)
(Revised August 3 2005)
(Accepted August 4 2005)
(October 3 2005)


Key Words: parasitic protozoa; trypanosomatids; Leishmania (Viannia) braziliensis; leishmaniasis; proteases; metalloproteases.

Correspondence:
c1 Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Av. Brasil 4365, Manguinhos, CEP: 21045-900, Rio de Janeiro, RJ, Brazil. Tel: +55 21 3865 8153. Fax: +55 21 2590 3495. E-mail: jbj@ioc.fiocruz.br


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