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Proteolytic expression in Blastocrithidia culicis: influence of the endosymbiont and similarities with virulence factors of pathogenic trypanosomatids

  • C. M. D'AVILA-LEVY (a1), F. M. ARAUJO (a1), A. B. VERMELHO (a1), R. M. A. SOARES (a1), A. L. S. SANTOS (a1) and M. H. BRANQUINHA (a1)...

Blastocrithidia culicis is an insect trypanosomatid that presents bacterial endosymbionts. The cell-associated and secreted proteinases of the endosymbiont-bearing and aposymbiotic strains were compared through the incorporation of proteinaceous substrates into sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Few qualitative changes could be detected in the proteolytic zymograms in the 2 strains studied when gelatin, casein, haemoglobin or bovine serum albumin (BSA) were tested. However, the level of proteolytic activities was significantly higher in the aposymbiotic strain. Some of the B. culicis proteins reacted in Western blots with antibodies raised against gp63, a zinc-metalloproteinase, and cruzipain, a cysteinyl-proteinase, which are virulence factors of the human pathogenic trypanosomatids, Leishmania spp. and Trypanosoma cruzi, respectively. The anti-cross-reacting determinant (CRD) antibody recognized 2 polypeptides (50 and 58 kDa) in the spent culture media and in the supernatant from glycosylphosphatidylinositol-phospholipase C (GPI-PLC)-treated cells, suggesting that these proteins are GPI-anchored to the plasma membrane. In addition, the anti-gp63 reacted with the 50 kDa protein. The identification of protein homologues in trypanosomatids with distinct life-cycles may help to determine the importance of proteinases in trypanosomatids.

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BRANQUINHA, M. H., VERMELHO, A. B., GOLDENBERG, S. & BONALDO, M. C. ( 1996). Ubiquity of cysteine- and metalloproteinase activities in a wide range of trypanosomatids. Journal of Eukaryotic Microbiology43, 131135.

CAZZULO, J. J., STOKA, V. & TURK, V. ( 2001). The major cysteine proteinase of Trypanosoma cruzi: a valid target for chemotherapy of Chagas disease. Current Pharmaceutical Design7, 11431156.

CUEVAS, I. C., CAZZULO, J. J. & SÁNCHEZ, D. O. ( 2003). gp63 homolgues in Trypanosoma cruzi: surface antigens with metalloprotease activity and a possible role in host cell infection. Infection and Immunity71, 57395749.

D'AVILA-LEVY, C. M., MELO, A. C. N., VERMELHO, A. B. & BRANQUINHA, M. H. ( 2001). Differential expression of proteolytic enzymes in endosymbiont-harboring Crithidia species. FEMS Microbiology Letters202, 7377.

D'AVILA-LEVY, C. M., SOUZA, R. F., GOMES, R. C., VERMELHO, A. B. & BRANQUINHA, M. H. ( 2003a). A metalloproteinase extracellularly released by Crithidia deanei. Canadian Journal of Microbiology49, 625632.

D'AVILA-LEVY, C. M., SOUZA, R. F., GOMES, R. C., VERMELHO, A. B. & BRANQUINHA, M. H. ( 2003b). A novel extracellular calcium-dependent cysteine proteinase from Crithidia deanei. Archives of Biochemistry and Biophysics420, 18.

DE SOUZA, W. & MOTTA, M. C. M. ( 1999). Endosymbiosis in protozoa of the Trypanosomatidae family. FEMS Microbiology Letters173, 18.

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

GREIG, S. & ASHALL, F. ( 1990). Electrophoretic detection of Trypanosoma cruzi peptidases. Molecular and Biochemical Parasitology39, 3138.

GRUSZYNSK, A. E., DEMASTER, A., HOOPER, N. M. & BANGS, J. D. ( 2003). Surface coat remodeling during differentiation of Trypanosoma brucei. The Journal of Biological Chemistry278, 2466524672.

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

JONES, B. L., FONTANINI, D., JARVINEN, D. & PEKKARINEN, A. ( 1998). Simplified endoproteinase assays using gelatin or azogelatin. Analytical Biochemistry263, 214220.

LACOUNT, D. J., GRUSZYNSKI, A. E., GRANDGNETT, P. M., BANGS, J. D. & DONELSON, J. E. ( 2003). Expression and function of the Trypanosoma brucei major surface protease (GP63) genes. The Journal of Biological Chemistry278, 2465824664.

MELO, A. C. N., GIOVANNI-DE-SIMMONE, S., BRANQUINHA, M. H. & VERMELHO, A. B. ( 2001). Crithidia guilhermei: purification and partial characterization of a 62-kDa extracellular metalloproteinase. Experimental Parasitology97, 18.

MELO, A. C. N., D'AVILA-LEVY, C. M., BRANQUINHA, M. H. & VERMELHO, A. B. ( 2002). Crithidia guilhermei: gelatin- and hemoglobin-degrading extracellular metalloproteinases. Experimental Parasitology102, 150156.

ROITMAN, C., ROITMAN, I. & AZEVEDO, H. P. ( 1972). Growth of an insect trypanosomatid at 37 °C in a defined medium. Journal of Protozoology19, 346349.

SAJID, M. & McKERROW, J. H. ( 2002). Cysteine proteases of parasitic organisms. Molecular and Biochemical Parasitology120, 121.

SANTOS, A. L. S., FERREIRA, A., FRANCO, A. V., ALVIANO, C. S. & SOARES, R. M. A. ( 1999). Characterization of proteinases in Herpetomonas anglusteri and Herpetomonas roitmani. Current Microbiology39, 6164.

SANTOS, A. L. S., ABREU, C. M., BATISTA, L. M., ALVIANO, C. S. & SOARES, R. M. A. ( 2001). Cell-associated and extracellular proteinases in Blastocrithidia culicis: influence of growth conditions. Current Microbiology43, 100106.

SANTOS, A. L. S., ABREU, C. M., ALVIANO, C. S. & SOARES, R. M. A. ( 2002). Activation of the glycosylphosphatidylinositol-anchored membrane proteinases upon release from Herpetomonas samuelpessoai by phospholipase C. Current Microbiology45, 293298.

SCHNEIDER, P. & GLASER, T. A. ( 1993). Characterization of a surface metalloprotease from Herpetomonas samuelpessoai and comparison with Leishmania major promastigote surface protease. Molecular and Biochemical Parasitology58, 277282.

SOARES, R. M. A., SANTOS, A. L. S., BONALDO, M. C., ANDRADE, A. F. B., ALVIANO, C. S., ANGLUSTER, J. & GOLDENBERG, S. ( 2003). Leishmania (Leishmania) amazonensis: differential expression of proteinases and cell-surface polypeptides in avirulent and virulent promastigotes. Experimental Parasitology104, 104112.

SOUSA, M. A. ( 1994). Trypomastigotes in cultures of Blastocrithidia culicis (Novy, MacNeal & Torrey, 1907) (Kinetoplastida: Trypanosomatidae). Memórias do Instituto Oswaldo Cruz89, 111112.

VERMELHO, A. B., ALMEIDA, F. V. S., BRONZATO, L. S. & BRANQUINHA, M. H. ( 2003). Extracellular metalloproteinases in Phytomonas serpens. Canadian Journal of Microbiology49, 221224.

YAO, C., DONELSON, J. E. & WILSON, M. E. ( 2003). The major surface protease (MSP or GP63) of Leishmania sp. Biosynthesis, regulation of expression, and function. Molecular and Biochemical Parasitology132, 116.

WALLACE, F. G. ( 1966). The trypanpanosomatid parasites of insects and arachnids. Experimental Parasitology18, 124193.

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