Hostname: page-component-cd9895bd7-gvvz8 Total loading time: 0 Render date: 2024-12-26T22:48:40.427Z Has data issue: false hasContentIssue false
  • English
  • Français

Use of proteolytic enzymes as an additional tool for trypanosomatid identification

Published online by Cambridge University Press:  13 December 2004

A. L. S. SANTOS ,
C. M. ABREU ,
C. S. ALVIANO  and
R. M. A. SOARES
A. L. S. SANTOS
Affiliation:
Laboratório de Biologia de Protistas, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Av. Brigadeiro Trompowsky, sem número, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil
C. M. ABREU
Affiliation:
Laboratório de Biologia de Protistas, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Av. Brigadeiro Trompowsky, sem número, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil
C. S. ALVIANO
Affiliation:
Laboratório de Biologia de Protistas, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Av. Brigadeiro Trompowsky, sem número, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil
R. M. A. SOARES
Affiliation:
Laboratório de Biologia de Protistas, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes (IMPPG), Centro de Ciências da Saúde (CCS), Universidade Federal do Rio de Janeiro (UFRJ), Av. Brigadeiro Trompowsky, sem número, Ilha do Fundão, Rio de Janeiro, RJ 21941-590, Brazil
Get access Rights & Permissions [Opens in a new window]

Abstract

The expression of proteolytic activities in the Trypanosomatidae family was explored as a potential marker to discriminate between the morphologically indistinguishable flagellates isolated from insects and plants. We have comparatively analysed the proteolytic profiles of 19 monoxenous trypanosomatids (Herpetomonas anglusteri, H. samuelpessoai, H. mariadeanei, H. roitmani, H. muscarum ingenoplastis, H. muscarum muscarum, H. megaseliae, H. dendoderi, Herpetomoas sp., Crithidia oncopelti, C. deanei, C. acanthocephali, C. harmosa, C. fasciculata, C. guilhermei, C. luciliae, Blastocrithidia culicis, Leptomonas samueli and Lept. seymouri) and 4 heteroxenous flagellates (Phytomonas serpens, P. mcgheei, Trypanosoma cruzi and Leishmania amazonensis) by in situ detection of enzyme activities on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS–PAGE ) containing co-polymerized gelatine as substrate, in association with specific proteinase inhibitors. All 23 trypanosomatids expressed at least 1 acidic proteolytic enzyme. In addition, a characteristic and specific pattern of cell-associated metallo and/or cysteine proteinases was observed, except for the similar profiles detected in 2 Herpetomonas (H. anglusteri and H. samuelpessoai) and 3 Crithidia (C. fasciculata, C. guilhermei and C. luciliae) species. However, these flagellates released distinct secretory proteinase profiles into the extracellular medium. These findings strongly suggest that the association of cellular and secretory proteinase pattern could represent a useful marker to help trypanosomatid identification.

Keywords

cell-associated proteinasessecretory proteinasescysteine proteinasesgelatine-polyacrylamide gel electrophoresismetalloproteinasesTrypanosomatidae family
Type
Research Article
Information
Parasitology , Volume 130 , Issue 1 , January 2005 , pp. 79 - 88
Copyright
© 2004 Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

ABREU FILHO, B. A., DIAS FILHO, B. P., VERMELHO, A. B., JANKEVICIUS, S. I., JANKEVICIUS, J. V. & SANTOS, R. L. ( 2001). Surface component characterization as taxonomic tools for Phytomonas spp identification. Parasitology Research 87, 138–144.CrossRefGoogle Scholar
ALFIERI, S. C., BALANCO, J. M. F. & PRAL, E. M. F. ( 1995). A radioiodinated peptidyl diazomethane detects similar cysteine proteinases in amastigotes and promastigotes of Leishmania (L.) mexicana and L. (L.) amazonensis. Parasitology Research 81, 240–244.Google Scholar
BARRETT, A. J., RAWLINGS, N. D. & O'BRIEN, E. A. ( 2001). The MEROPS database as a protease information system. Journal of Structural Biology 134, 95–102.CrossRefGoogle Scholar
BOISSEAU-GARSAUD, A. M., CALES-QUIST, D., DESBOIS, N., JOUANNELLE, J., JOUANNELLE, A., PRATLONG, F. & DEDET, J. P. ( 2000). A new case of cutaneous infections by a presumed monoxenous trypanosomatid in the island of Martinique (French West Indies). Transactions of the Royal Society of Tropical Medicine and Hygiene 94, 51–52.CrossRefGoogle Scholar
BONALDO, M. C., D'ESCOFFIER, L. N., SALLES, J. M. & GOLDENBERG, S. ( 1991). Characterization and expression of protease during Trypanosoma cruzi metacyclogenesis. Experimental Parasitology 73, 44–51.CrossRefGoogle Scholar
BRANDÃO, A. A., MIRANDA, A., DEGRAVE, W. M. & SOUSA, M. A. ( 2000). The heterogeneity of choanomastigote-shaped trypanosomatids as analyzed by their kDNA minicircle size: taxonomic implications. Parasitology Research 86, 809–812.CrossRefGoogle Scholar
BRANQUINHA, M. H., BARRETO-BERGTER, E., MEIRELLES, M. N. L. & VERMELHO, A. B. ( 1994). Glycolipid and protein profiles in trypanosomatids. Parasitology Research 80, 336–341.CrossRefGoogle Scholar
BRANQUINHA, M. H., VERMELHO, A. B., GOLDENBERG, S. & BONALDO, M. C. ( 1996). Ubiquity of cysteine- and metalloproteinase in a wide range of trypanosomatids. Journal of Eukaryotic Microbiology 43, 131–135.CrossRefGoogle Scholar
CAMARGO, E. P., KASTELEIN, P. & ROITMAN, I. ( 1990). Trypanosomatid parasites of plants (Phytomonas). Parasitology Today 6, 22–25.CrossRefGoogle Scholar
CAMARGO, E. P., SBRAVATE, C., TEIXEIRA, M. M. G., ULIANA, S. R. B., SOARES, M. B. M., AFFONSO, H. T. & FLOETER-WINTER, L. ( 1992). Ribosomal DNA restriction analysis and synthetic oligonucleotide probing in the identification of genera of lower trypanosomatids. Journal of Parasitology 78, 40–48.CrossRefGoogle Scholar
D'ÁVILA-LEVY, C. M., ARAÚJO, F. M., VERMELHO, A. B., BRANQUINHA, M. H., ALVIANO, C. S., SOARES, R. M. A. & SANTOS, A. L. S. ( 2004). Distribution of glycoproteins recognized by different lectins in seven choanomastigote-shaped trypanosomatids: taxonomic implications. FEMS Microbiology Letters 231, 171–176.CrossRefGoogle Scholar
D'ÁVILA-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 Letters 202, 73–77.CrossRefGoogle Scholar
DEDET, J. P. & PRATLONG, F. ( 2000). Leishmania, Trypanosoma and monoxenous trypanosomatids as emerging opportunistic agents. Journal of Eukaryotic Microbiology 47, 37–39.CrossRefGoogle Scholar
DEDET, J. P., ROCHE, B., PRATLONG, F., CALES-QUIST, D., JOUANNELLE, J., BENICHOU, J. C. & HUERRE, M. ( 1995). Diffuse cutaneous infection caused by a presumed monoxenous trypanosomatid in a patient infected with HIV. Transactions of the Royal Society of Tropical Medicine and Hygiene 89, 644–646.CrossRefGoogle Scholar
ETGES, R. ( 1992). Identification of a surface metalloproteinase on 13 species of Leishmania isolated from humans, Crithidia fasciculata and Herpetomonas samuelpessoai. Acta Tropica 50, 205–217.CrossRefGoogle Scholar
FERNANDES, O., TEIXEIRA, M. M. G., STURM, N. R., SOUSA, M. A., CAMARGO, E. P., DEGRAVE, W. M. & CAMPBELL, D. A. ( 1997). Mini-exon gene sequences define six groups within the genus Crithidia. Journal of Eukaryotic Microbiology 44, 535–539.CrossRefGoogle Scholar
FRANCO, A. M. R., MOMEN, H., NAIFF, R. D., MOREIRA, C. F. S., DEANE, M. P. & GRIMALDI, G. ( 1996). Enzyme polymorphism in Endotrypanum and numerical analysis of isoenzyme data. Parasitology 113, 39–48.CrossRefGoogle Scholar
HEUSSEN, C. & DOWDLE, E. B. ( 1980). Electrophoretic analysis of plasminogen activators in polyacrilamide gels containing sodium dodecyl sulphate and copolymerized substrates. Analytical Biochemistry 102, 196–202.CrossRefGoogle Scholar
JAFFE, C. L. & DWYER, D. M. ( 2003). Extracellular release of the surface metalloprotease, gp63, from Leishmania and insect trypanosomatids. Parasitology Research 91, 229–237.CrossRefGoogle Scholar
JIMÉNEZ, M. I., LÓPEZ-VÉLEZ, R., MOLINA, R., CAÑAVATE, C. & ALVAR, J. ( 1996). HIV co-infection with a currently non-pathogenic flagellate. Lancet 347, 264–265.CrossRefGoogle Scholar
LWOFF, M. & LWOFF, A. ( 1931). Recherches sur la morphologie de Leptomonas oncopelti Noguchi et Tilden et Leptomonas fasciculata Novy, Mac Neal et Torrey. Arquives Zoologic Experimental Géneral 71, 21–37.Google Scholar
MARCHÉ, S., ROTH, C., PHILIPPE, H., DOLLET, M. & BALTZ, T. ( 1995). Characterization and detection of plant trypanosomatids by sequence analysis of the small subunit ribosomal RNA gene. Molecular and Biochemical Parasitology 71, 15–26.CrossRefGoogle Scholar
McGHEE, R. B. & COSGROVE, W. B. ( 1980). Biology and physiology of the lower trypanosomatids. Microbiology Reviews 44, 140–173.Google Scholar
McKERROW, J. H., SUN, E., ROSENTHAL, P. J. & BOUVIER, J. ( 1993). The proteases and pathogenicity of parasitic protozoa. Annual Review of Microbiology 47, 821–853.CrossRefGoogle Scholar
MEDINA-ACOSTA, E., FRANCO, A. M. R., JANSEN, A. M., SAMPOL, M., NEVÉS, N., PONTES-DE-CARVALHO, L., GRIMALDI, G. & NUSSENZWEIG, V. ( 1994). Trans-sialidase and sialidase activities discriminate between morphologically indistinguishable trypanosomatids. European Journal of Biochemistry 225, 333–339.CrossRefGoogle Scholar
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 Parasitology 97, 1–8.Google Scholar
MICHAUD, D., CANTIN, L., RAWORTH, D. A. & VRAIN, T. C. ( 1996). Assessing the stability of cystatin/cysteine proteinase complexes using mildly-denaturing gelatine-polyacrylamide gel electrophoresis. Electrophoresis 17, 74–79.CrossRefGoogle Scholar
MILLER, R. F. ( 2000). Clinical presentation and significance of emerging opportunistic infections. Journal of Eukaryotic Microbiology 47, 21–23.CrossRefGoogle Scholar
MOMEN, H. ( 2001). Some current problems in the systematics of trypanosomatids. International Journal for Parasitology 31, 640–642.CrossRefGoogle Scholar
MOMEN, H. ( 2002). Molecular taxonomy of trypanosomatids: some problems and pitfalls. Archives of Medical Research 33, 413–415.CrossRefGoogle Scholar
NOYES, H., PRATLONG, F., CHANCE, M., ELLIS, J., LANOTTE, G. & DEDET, J. P. ( 2002). A previously unclassified trypanosomatid responsible for human cutaneous lesions in Martinique (French West Indies) is the most divergent member of the genus Leishmania ss. Parasitology 124, 17–24.CrossRefGoogle Scholar
PACHECO, R. S., MARZOCHI, M. C. A., PIRES, M. Q., BRITO, C. M. M., MADEIRA, M. F. & BARBOSA-SANTOS, E. G. O. ( 1998). Parasite genotypically related to a monoxenous trypanosomatid of dog's flea causing opportunistic infection in an HIV positive patient. Memórias do Instituto Oswaldo Cruz 93, 531–537.CrossRefGoogle Scholar
PODLIPAEV, S. ( 2001). The more insect trypanosomatids under study-the more diverse Trypanosomatidae appears. International Journal for Parasitology 31, 648–652.CrossRefGoogle Scholar
RAO, M. B., TANKSALE, A. M., GHATGE, M. S. & DESHPANDE, V. V. ( 1998). Molecular and biotechnological aspects of microbial proteases. Microbiology and Molecular Biology Reviews 62, 597–635.Google Scholar
ROGERS, W. E. & WALLACE, F. G. ( 1971). Two new subspecies of Herpetomonas muscarum (Leidy 1856) Kent 1880. Journal of Protozoology 18, 645–649.CrossRefGoogle Scholar
SAJID, M. & McKERROW, J. H. ( 2002). Cysteine proteases of parasitic organisms. Molecular and Biochemical Parasitology 120, 1–21.CrossRefGoogle Scholar
SANTOS, A. L. S., ABREU, C. M., ALVIANO, C. S. & SOARES, R. M. A. ( 2002 a). Activation of the glycosylphosphatidylinositol membrane proteinase upon released from Herpetomonas samuelpessoai by phospholipase C. Current Microbiology 45, 293–298.Google Scholar
SANTOS, A. L. S., ALVIANO, C. S. & SOARES, R. M. A. ( 2002 b). Detection of sialoglycomolecules in five plant trypanosomatids and in an insect phytophagous isolate. FEMS Microbiology Letters 214, 19–23.Google Scholar
SANTOS, A. L. S., FERREIRA, A., FRANCO, V. A., ALVIANO, C. S. & SOARES, R. M. A. ( 1999). Characterization of proteinases in Herpetomonas anglusteri and Herpetomonas roitmani. Current Microbiology 39, 61–64.CrossRefGoogle Scholar
SANTOS, A. L. S., RODRIGUES, M. L., ALVIANO, C. S., ANGLUSTER, J. & SOARES, R M. A. ( 2003). Herpetomonas samuelpessoai: dimethylsulfoxide-induced differentiation is influenced by proteinase expression. Current Microbiology 46, 11–17.CrossRefGoogle Scholar
SANTOS, A. L. S., SOUTO-PADRÓN, T., ALVIANO, C. S., LOPES, A. H. C. S., SOARES, R. M. A. & MEYER-FERNANDES, J. R. ( 2002 c). Secreted phosphatase activity induced by dimethylsulfoxide in Herpetomonas samuelpessoai. Archives of Biochemistry and Biophysics 405, 191–198.Google Scholar
SILVA, P. M. F. E., SOLÉ-CAVA, A. M., SOARES, M. J., MOTTA, M. C. M., FIORINI, J. E. & DE SOUZA, W. ( 1991). Herpetomonas roitmani (Fiorini et al., 1989) n. comb.: a trypanosomatid with a bacterium-like endosymbiont in the cytoplasm. Journal of Protozoology 38, 489–494.Google Scholar
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 Parasitology 104, 104–112.CrossRefGoogle Scholar
SOUSA, M. A. & CÔRTE-REAL, S. ( 1991). Postnuclear kinetoplast in choanomastigotes of Crithidia deanei Carvalho, 1973: proposal of a new genus. Revista do Instituto de Medicina Tropical de São Paulo 33, S8.Google Scholar
TEIXEIRA, M. M. G., CAMPANER, M. & CAMARGO, E. P. ( 1995). Characterization of the target antigens of Phytomonas-specific monoclonal antibodies. Journal of Eukaryotic Microbiology 42, 232–237.CrossRefGoogle Scholar
TEIXEIRA, M. M. G., TAKATA, C. S. A., CONCHON, I., CAMPANER, M. & CAMARGO, E. P. ( 1997). Ribosomal and kDNA markers distinguish two subgroups of Herpetomonas among old species and new trypanosomatids isolated from flies. Journal of Parasitology 83, 58–65.CrossRefGoogle Scholar
VICKERMAN, K. ( 1994). The evolutionary expansion of the trypanosomatid flagellates. International Journal for Parasitology 24, 1317–1331.CrossRefGoogle Scholar
WALLACE, F. G. ( 1966). The trypanosomatid parasites of insects and arachnids. Experimental Parasitology 18, 124–193.CrossRefGoogle Scholar
WALLACE, F. G., CAMARGO, E. P., McGHEE, R. B. & ROITMAN, I. ( 1983). Guidelines for the description of new species of lower trypanosomatids. Journal of Protozoology 30, 308–313.CrossRefGoogle Scholar
WALLACE, F. G., WAGNER, M. & ROGERS, W. E. ( 1973). Varying kinetoplast ultrastructure in two species of Herpetomonas muscarum. Journal of Protozoology 20, 218–222.CrossRefGoogle Scholar