Heterogeneous production of metallo-type peptidases in parasites belonging to the family Trypanosomatidae.
Proteolytic enzymes play a central role in the physiology of all living organisms, participating in several metabolic pathways and in different phases of parasite-host interactions. We have identified cell-associated peptidase activities in 33 distinct flagellates, including representatives of almost all known trypanosomatid genera parasitizing insects (Herpetomonas, Crithidia, Leishmania, Trypanosoma, Leptomonas, Phytomonas, Blastocrithidia and Endotrypanum) as well as the biflagellate kinetoplastid Bodo, by using SDS-PAGE containing gelatin as co-polymerized substrate and proteolytic inhibitors. Under the alkaline pH (9.0) conditions employed, all the flagellates presented at least one peptidase, with the exception of Crithidia acanthocephali and Phytomonas serpens, which did not display any detectable proteolytic enzyme activity. All the proteolytic activities were completely inhibited by 1,10-phenanthroline, a zinc-chelating agent, putatively identifying these activities as metallo-type peptidases. EDTA and EGTA, two other metallopeptidase inhibitors, E-64 (a cysteine peptidase inhibitor), pepstatin A (an aspartyl peptidase inhibitor) and PMSF (a serine peptidase inhibitor) did not interfere with the metallopeptidase activities detected in the studied trypanosomatids. Conversely, Bodo-derived peptidases were resistant to 1,10-phenanthroline and only partially inhibited by EDTA, showing a distinct inhibition profile. Together, our data demonstrated great heterogeneity of expression of metallopeptidases in a wide range of parasites belonging to the family Trypanosomatidae.
dos Santos
André Luis Souza
AL
Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, Cidade Universitária, Ilha do Fundao, 21941-902 Rio de Janeiro, RJ, Brazil. andre@micro.ufrj.br
Soares
Rosangela Maria de Araújo
RM
Alviano
Celuta Sales
CS
Kneipp
Lucimar Ferreira
LF
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005780:Gelatin / Q000378:metabolism; D007313:Insecta / Q000469:parasitology; D008857:Microscopy, Interference; D010447:Peptide Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D013045:Species Specificity; D014351:Trypanosomatina / Q000201:enzymology*
D005780:Gelatin; D010447:Peptide Hydrolases