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Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis

Published online by Cambridge University Press:  21 October 2005

V. G. DUSCHAK ,
M. BARBOZA ,
G. A. GARCÍA ,
E. M. LAMMEL ,
A. S. COUTO  and
E. L. D. ISOLA
V. G. DUSCHAK
Affiliation:
Instituto Nacional de Parasitología ‘Dr Mario Fatala Chabén’, ANLIS-Malbrán, Ministerio de Salud y Ambiente, Av. Paseo Colon 568, 1063 Buenos Aires, Argentina
M. BARBOZA
Affiliation:
Instituto de Investigaciones Biotecnológicas–INTECH, Universidad Nacional de General San Martín, 1650 Buenos Aires, Argentina
G. A. GARCÍA
Affiliation:
Instituto Nacional de Parasitología ‘Dr Mario Fatala Chabén’, ANLIS-Malbrán, Ministerio de Salud y Ambiente, Av. Paseo Colon 568, 1063 Buenos Aires, Argentina
E. M. LAMMEL
Affiliation:
Departamento de Microbiología, Parasitología e Inmunología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, Buenos Aires, Argentina
A. S. COUTO
Affiliation:
CIHIDECAR, Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Pabellón 2, Ciudad Universitaria, 1428 Buenos Aires, Argentina
E. L. D. ISOLA
Affiliation:
Departamento de Microbiología, Parasitología e Inmunología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, Buenos Aires, Argentina
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Abstract

With the aim to study proteinases released to the culture medium during Trypanosoma cruzi metacyclogenesis, the presence of cysteine proteinases (CPs) was analysed in culture supernatants obtained throughout the differentiation induced by stimulation of epimastigotes with Triatoma infestans hindgut homogenate. In SDS-gelatin containing gels, an important endopeptidase activity with apparent molecular weight range between 97 and 116 kDa was encountered at pH 6, which was abolished by the specific cysteine proteinase inhibitor E-64 and TLCK, but not by pepstatin, 1,10 phenantroline or PMSF. This novel CP, named TcCPmet, showed affinity to cystatin-Sepharose, denoting its thiol-proteinase character as well as to ConA-Sepharose, indicating it contains N-linked oligosaccharides. However, it presented a different elution pattern on ConA-Sepharose than cruzipain and, in addition, it was not recognized by anti-cruzipain serum, facts that strongly suggest the different nature of both CPs. Moroever, evidence is presented indicating that TcCPmet was able to hydrolyse the same chromogenic peptides as cruzipain at optimal alkaline pH values, although with a different order of effectiveness. Our results indicate the presence of a novel CP secreted by metacyclic trypomastigotes and reinforces the important role of these enzymes in metacyclogenesis.

Keywords

Trypanosoma cruzicysteine proteinasescruzipainmetacyclogenesis
Type
Research Article
Information
Parasitology , Volume 132 , Issue 3 , March 2006 , pp. 345 - 355
Copyright
© 2005 Cambridge University Press

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