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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
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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

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