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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)...
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.

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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: jbj@ioc.fiocruz.br
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REFERENCES

Alexander, J., Coombs, G. H. and Mottram, J. C. ( 1998). Leishmania mexicana cysteine proteinase-deficient mutants have attenuated virulence for mice and potentiate a Th1 response. Journal of Immunology 161, 67946801.
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 Parasitology 109, 3848.
Bates, P. A., Hermes, I. and Dwyer, D. M. ( 1989). Leishmania donovani: immunochemical localization and secretory mechanism of soluble acid phosphatase. Experimental Parasitology 68, 335346.
Beynon, R. and Bond, J. S. ( 2001). Proteolytic Enzymes, 2nd Edn. Oxford University Press, New York.
Blum, J. J. and Opperdoes, F. R. ( 1994). Secretion of sucrase by Leishmania donovani. Journal of Eukaryotic Microbiology 41, 228231.
Bontempi, E. and Cazzulo, J. J. ( 1990). Digestion of human immunoglobulin G by the major cysteine proteinase (cruzipain) from Trypanosoma cruzi. FEMS Microbiology Letters 58, 337341.
Bouvier, J., Etges, R. J. and Bordier, C. ( 1985). Identification and purification of membrane and soluble forms of the major surface protein of Leishmania promastigotes. The Journal of Biological Chemistry 260, 1550415509.
Brittingham, A., Morrison, C. J., McMaster, W. R., McGwire, B. S., Chang, K. P. and Mosser, D. M. ( 1995). Role of the Leishmania surface protease gp63 in complement fixation, cell adhesion, and resistance to complement-mediated lysis. Journal of Immunology 155, 31023111.
Brittingham, A., Chen, G., McGwire, B. S., Chang, K. P. and Mosser, D. M. ( 1999). Interaction of Leishmania gp63 with cellular receptors for fibronectin. Infection and Immunity 67, 44774484.
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 Immunology 171, 37113717.
Canto-Lara, S. B., Cardenas-Maruffo, M. F., Vargas-Gonzalez, A. and Andrade-Narvaez, F. ( 1998). Isoenzyme characterization of Leishmania isolated from human cases with localized cutaneous leishmaniasis from the State of Campeche, Yucatan Peninsula, Mexico. American Journal of Tropical Medicine and Hygiene 58, 444447.
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, USA 83, 100104.
Chaudhuri, G., Chaudhuri, M., Pan, A. and Chang, K. P. ( 1989). Surface acid proteinase (gp63) of Leishmania mexicana. A metalloenzyme capable of protecting liposome-encapsulated proteins from phagolysosomal degradation by macrophages. The Journal of Biological Chemistry 264, 74837489.
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 Medicine 162, 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 Medicine 201, 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 Microbiology 41, 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 Parasitology 104, 104112.
Etges, R. ( 1992). Identification of a surface metalloproteinase on 13 species of Leishmania isolated from humans, Crithidia fasciculata, and Herpetomonas samuelpessoai. Acta Tropica 50, 205217.
Etges, R., Bouvier, J. and Bordier, C. ( 1986). The major surface protein of Leishmania promastigotes is a protease. The Journal of Biological Chemistry 261, 90989101.
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 Parasitology 14, 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, USA 78, 76247628.
Gardiner, P. R., Jaffe, C. L. and Dwyer, D. M. ( 1984). Identification of cross-reactive promastigote cell surface antigens of some leishmanial stocks by 125I labeling and immunoprecipitation. Infection and Immunity 43, 637643.
Grimaldi, G. Jr. and Tesh, R. B. ( 1993). Leishmaniases of the New World: current concepts and implications for future research. Clinical Microbiology Reviews 6, 230250.
Grimaldi, G. Jr., Tesh, R. B. and McMahon-Pratt, D. ( 1989). A review of the geographic distribution and epidemiology of leishmaniasis in the New World. American Journal of Tropical Medicine and Hygiene 41, 687725.
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 Pasteur 6, 646649.
Jacobson, R. L. and Schlein, Y. ( 1997). Cellulase activity of Leishmania major in the sandfly vector and in culture. Journal of Eukaryotic Microbiology 44, 216219.
Jaffe, C. L. and Dwyer, D. M. ( 2003). Extracellular release of the surface metalloprotease, gp63, from Leishmania and insect trypanosomatids. Parasitology Research 91, 229237.
Klemba, M. and Goldberg, D. E. ( 2002). Biological roles of proteases in parasitic protozoa. Annual Review of Biochemistry 71, 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 Tropica 56, 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, USA 89, 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 Parasitology 24, 8995.
Mahmoudzadeh-Niknam, H. and McKerrow, J. H. ( 2004). Leishmania tropica: cysteine proteases are essential for growth and pathogenicity. Experimental Parasitology 106, 158163.
Marsden, P. D. ( 1986). Mucosal leishmaniasis (“espundia” Escomel, 1911). Transactions of the Royal Society of Tropical Medicine and Hygiene 80, 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 Chemistry 277, 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 Immunity 71, 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 Parasitology 57, 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 Parasitology 37, 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 Parasitology 105, 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 Microbiology 1, 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, USA 93, 60086013.
North, M. J. and Coombs, G. H. ( 1981). Proteinases of Leishmania mexicana amastigotes and promastigotes: analysis by gel electrophoresis. Molecular and Biochemical Parasitology 3, 293300.
Pupkis, M. F. and Coombs, G. H. ( 1984). Purification and characterization of proteolytic enzymes of Leishmania mexicana mexicana amastigotes and promastigotes. Journal of General Microbiology 130, 23752383.
Que, X. and Reed, S. L. ( 2000). Cysteine proteinases and the pathogenesis of amebiasis. Clinical Microbiology Reviews 13, 196206.
Robertson, C. D. and Coombs, G. H. ( 1992). Stage-specific proteinases of Leishmania mexicana mexicana promastigotes. FEMS Microbiology Letters 73, 127132.
Rosenthal, P. J. ( 1999). Proteases of protozoan parasites. Advances in Parasitology 43, 105159.
Russell, D. G. and Wilhelm, H. ( 1986). The involvement of the major surface glycoprotein (gp63) of Leishmania promastigotes in attachment to macrophages. Journal of Immunology 136, 26132620.
Sajid, M. and McKerrow, J. H. ( 2002). Cysteine proteases of parasitic organisms. Molecular and Biochemical Parasitology 120, 121.
Saravia, N. G., Holguin, A. F., McMahon-Pratt, D. and D'Alessandro, A. ( 1985). Mucocutaneous leishmaniasis in Colombia: Leishmania braziliensis subspecies diversity. American Journal of Tropical Medicine and Hygiene 34, 714720.
Schlein, Y. ( 1993). Leishmania and sandflies: interactions in the life-cycle and transmission. Parasitology Today 9, 255258.
Schneider, P. and Glaser, T. A. ( 1993). Characterization of two soluble metalloexopeptidases in the protozoan parasite Leishmania major. Molecular and Biochemical Parasitology 62, 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 Research 93, 328331.
Weigle, K. A., Saravia, N. G., de Davalos, M., Moreno, L. H. and D'Alessandro, A. ( 1986). Leishmania braziliensis from the Pacific coast region of Colombia: foci of transmission, clinical spectrum and isoenzyme phenotypes. American Journal of Tropical Medicine and Hygiene 35, 722731.
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 Parasitology 121, 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 Entomology 24, 587589.
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