Skip to main content
    • Aa
    • Aa

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

  • P. CUERVO (a1) (a2) (a3), L. SABÓIA-VAHIA (a4) (a5), F. COSTA SILVA-FILHO (a5), O. FERNANDES (a1), E. CUPOLILLO (a3) and J. B. DE JESUS (a4) (a5)...

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.

Corresponding author
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:
Linked references
Hide All

This list contains references from the content that can be linked to their source. For a full set of references and notes please see the PDF or HTML where available.

Alves, C. R., Corte-Real, S., Bourguignon, S. C., Chaves, C. S. and Saraiva, E. M. ( 2005). Leishmania amazonensis: early proteinase activities during promastigote-amastigote differentiation in vitro. Experimental Parasitology109, 3848.

Bates, P. A., Hermes, I. and Dwyer, D. M. ( 1989). Leishmania donovani: immunochemical localization and secretory mechanism of soluble acid phosphatase. Experimental Parasitology68, 335346.

Blum, J. J. and Opperdoes, F. R. ( 1994). Secretion of sucrase by Leishmania donovani. Journal of Eukaryotic Microbiology41, 228231.

Buxbaum, L. U., Denise, H., Coombs, G. H., Alexander, J., Mottram, J. C. and Scott, P. ( 2003). Cysteine protease B of Leishmania mexicana inhibits host Th1 responses and protective immunity. Journal of Immunology171, 37113717.

Chang, C. S. and Chang, K. P. ( 1986). Monoclonal antibody affinity purification of a Leishmania membrane glycoprotein and its inhibition of leishmania-macrophage binding. Proceedings of the National Academy of Sciences, USA83, 100104.

Colomer-Gould, V., Glvao Quintao, L., Keithly, J. and Nogueira, N. ( 1985). A common major surface antigen on amastigotes and promastigotes of Leishmania species. The Journal of Experimental Medicine162, 902916.

Coombs, G. H. ( 1982). Proteinases of Leishmania mexicana and other flagellate protozoa. Parasitology 84, 149155.

Coppi, A., Pinzon-Ortiz, C., Hutter, C. and Sinnis, P. ( 2005). The Plasmodium circumsporozoite protein is proteolytically processed during cell invasion. The Journal of Experimental Medicine201, 2733.

Cupolillo, E., Brahim, L. R., Toaldo, C. B., de Oliveira-Neto, M. P., de Brito, M. E., Falqueto, A., de Farias Naiff, M. and Grimaldi, G.Jr. ( 2003). Genetic polymorphism and molecular epidemiology of Leishmania (Viannia) braziliensis from different hosts and geographic areas in Brazil. Journal of Clinical Microbiology41, 31263132.

De Araujo Soares, R. M., dos Santos, A. L., Bonaldo, M. C., de Andrade, A. F., Alviano, C. S., Angluster, J. and Goldenberg, S. ( 2003). Leishmania (Leishmania) amazonensis: differential expression of proteinases and cell-surface polypeptides in avirulent and virulent promastigotes. Experimental Parasitology104, 104112.

Etges, R. ( 1992). Identification of a surface metalloproteinase on 13 species of Leishmania isolated from humans, Crithidia fasciculata, and Herpetomonas samuelpessoai. Acta Tropica50, 205217.

Etges, R. J., Bouvier, J., Hoffman, R. and Bordier, C. ( 1985). Evidence that the major surface proteins of three Leishmania species are structurally related. Molecular and Biochemical Parasitology14, 141149.

Fong, D. and Chang, K. P. ( 1981). Tubulin biosynthesis in the developmental cycle of a parasitic protozoan, Leishmania mexicana: changes during differentiation of motile and nonmotile stages. Proceedings of the National Academy of Sciences, USA78, 76247628.

Grimaldi, G.Jr. and Tesh, R. B. ( 1993). Leishmaniases of the New World: current concepts and implications for future research. Clinical Microbiology Reviews6, 230250.

Hajmova, M., Chang, K. P., Kolli, B. and Volf, P. ( 2004). Down-regulation of gp63 in Leishmania amazonensis reduces its early development in Lutzomyia longipalpis. Microbes and Infection/Institut Pasteur6, 646649.

Jacobson, R. L. and Schlein, Y. ( 1997). Cellulase activity of Leishmania major in the sandfly vector and in culture. Journal of Eukaryotic Microbiology44, 216219.

Jaffe, C. L. and Dwyer, D. M. ( 2003). Extracellular release of the surface metalloprotease, gp63, from Leishmania and insect trypanosomatids. Parasitology Research91, 229237.

Klemba, M. and Goldberg, D. E. ( 2002). Biological roles of proteases in parasitic protozoa. Annual Review of Biochemistry71, 275305.

Leon, L. L., Temporal, R. M., Soares, M. J. and Grimaldi, G.Jr. ( 1994). Proteinase activities during temperature-induced stage differentiation of species complexes of Leishmania. Acta Tropica56, 289298.

Liu, X. and Chang, K. P. ( 1992). Extrachromosomal genetic complementation of surface metalloproteinase (gp63)-deficient Leishmania increases their binding to macrophages. Proceedings of the National Academy of Sciences, USA89, 49914995.

Lockwood, B. C., North, M. J., Scott, K. I., Bremner, A. F. and Coombs, G. H. ( 1987). The use of a highly sensitive electrophoretic method to compare the proteinases of trichomonads. Molecular and Biochemical Parasitology24, 8995.

Mahmoudzadeh-Niknam, H. and McKerrow, J. H. ( 2004). Leishmania tropica: cysteine proteases are essential for growth and pathogenicity. Experimental Parasitology106, 158163.

Marsden, P. D. ( 1986). Mucosal leishmaniasis (“espundia” Escomel, 1911). Transactions of the Royal Society of Tropical Medicine and Hygiene80, 859876.

McGrath, M. E., Eakin, A. E., Engel, J. C., McKerrow, J. H., Craik, C. S. and Fletterick, R. J. ( 1995). The crystal structure of cruzain: a therapeutic target for Chagas' disease. Journal of Molecular Biology 247, 251259.

McGwire, B. S., O'Connell, W. A., Chang, K. P. and Engman, D. M. ( 2002). Extracellular release of the glycosylphosphatidylinositol (GPI)-linked Leishmania surface metalloprotease, gp63, is independent of GPI phospholipolysis: implications for parasite virulence. The Journal of Biological Chemistry277, 88028809.

McGwire, B. S., Chang, K. P. and Engman, D. M. ( 2003). Migration through the extracellular matrix by the parasitic protozoan Leishmania is enhanced by surface metalloprotease gp63. Infection and Immunity71, 10081010.

Medina-Acosta, E., Karess, R. E. and Russell, D. G. ( 1993). Structurally distinct genes for the surface protease of Leishmania mexicana are developmentally regulated. Molecular and Biochemical Parasitology57, 3145.

Medina-Acosta, E., Karess, R. E., Schwartz, H. and Russell, D. G. ( 1989). The promastigote surface protease (gp63) of Leishmania is expressed but differentially processed and localized in the amastigote stage. Molecular and Biochemical Parasitology37, 263273.

Melo-Braga, M. B., da Rocha-Azevedo, B. and Silva-Filho, F. C. ( 2003). Tritrichomonas foetus: the role played by iron during parasite interaction with epithelial cells. Experimental Parasitology105, 111120.

Mottram, J. C., Brooks, D. R. and Coombs, G. H. ( 1998). Roles of cysteine proteinases of trypanosomes and Leishmania in host-parasite interactions. Current Opinion in Microbiology1, 455460.

Mottram, J. C., Souza, A. E., Hutchison, J. E., Carter, R., Frame, M. J. and Coombs, G. H. ( 1996). Evidence from disruption of the lmcpb gene array of Leishmania mexicana that cysteine proteinases are virulence factors. Proceedings of the National Academy of Sciences, USA93, 60086013.

North, M. J. and Coombs, G. H. ( 1981). Proteinases of Leishmania mexicana amastigotes and promastigotes: analysis by gel electrophoresis. Molecular and Biochemical Parasitology3, 293300.

Que, X. and Reed, S. L. ( 2000). Cysteine proteinases and the pathogenesis of amebiasis. Clinical Microbiology Reviews13, 196206.

Rosenthal, P. J. ( 1999). Proteases of protozoan parasites. Advances in Parasitology43, 105159.

Sajid, M. and McKerrow, J. H. ( 2002). Cysteine proteases of parasitic organisms. Molecular and Biochemical Parasitology120, 121.

Schlein, Y. ( 1993). Leishmania and sandflies: interactions in the life-cycle and transmission. Parasitology Today9, 255258.

Schneider, P. and Glaser, T. A. ( 1993). Characterization of two soluble metalloexopeptidases in the protozoan parasite Leishmania major. Molecular and Biochemical Parasitology62, 223231.

Silva-Lopez, R. E., Morgado-Diaz, J. A., Alves, C. R., Corte-Real, S. and Giovanni-De-Simone, S. ( 2004). Subcellular localization of an extracellular serine protease in Leishmania (Leishmania) amazonensis. Parasitology Research93, 328331.

Yao, C., Leidal, K. G., Brittingham, A., Tarr, D. E., Donelson, J. E. and Wilson, M. E. ( 2002). Biosynthesis of the major surface protease GP63 of Leishmania chagasi. Molecular and Biochemical Parasitology121, 119128.

Young, D. G., Morales, A., Kreutzer, R. D., Alexander, J. B., Corredor, A., Tesh, R. B., Ferro de Carrasquilla, C. and de Rodriguez, C. ( 1987). Isolation of Leishmania braziliensis (Kinetoplastida: Trypanosomatidae) from cryopreserved Colombian sand flies (Diptera: Psychodidae). Journal of Medical Entomology24, 587589.

Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

  • ISSN: 0031-1820
  • EISSN: 1469-8161
  • URL: /core/journals/parasitology
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



Full text views

Total number of HTML views: 2
Total number of PDF views: 7 *
Loading metrics...

Abstract views

Total abstract views: 57 *
Loading metrics...

* Views captured on Cambridge Core between September 2016 - 29th May 2017. This data will be updated every 24 hours.