Artículo

Duschak, V.G.; Barboza, M.; García, G.A.; Lammel, E.M.; Couto, A.S.; Isola, E.L.D. "Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis" (2006) Parasitology. 132(3):345-355
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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. © 2005 Cambridge University Press.

Registro:

Documento: Artículo
Título:Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis
Autor:Duschak, V.G.; Barboza, M.; García, G.A.; Lammel, E.M.; Couto, A.S.; Isola, E.L.D.
Filiación:Instituto Nacional de Parasitología 'Dr Mario Fatala Chabén', ANLIS-Malbrán, Ministerio de Salud Y Ambiente, Av. Paseo Colon 568, 1063 Buenos Aires, Argentina
Instituto de Investigaciones Biotecnológicas-INTECH, Universidad Nacional de General San Martín, 1650 Buenos Aires, Argentina
Departamento de Microbiología, Parasitología e Imnunología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, Buenos Aires, Argentina
CIHIDECAR, Departamento de Química Orgánica, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Departamento de Investigación, Instituto Nacional de Parasitología 'Dr Mario Fatala Chabén', Ministerio de Salud Y Ambiente, Av. Paseo Colón 568, 1063 Buenos Aires, Argentina
Palabras clave:Cruzipain; Cysteine proteinases; Metacyclogenesis; Trypanosoma cruzi; concanavalin A; cruzipain; cysteine proteinase; n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine; tosyllysyl chloromethyl ketone; animal tissue; article; binding affinity; controlled study; differentiation; enzyme activity; enzyme isolation; enzyme substrate complex; epimastigote; growth curve; life cycle; nonhuman; priority journal; Triatoma infestans; trypomastigote; Animals; Blotting, Western; Chromatography, Affinity; Cross Reactions; Culture Media; Culture Techniques; Cysteine Endopeptidases; Hydrogen-Ion Concentration; Life Cycle Stages; Protease Inhibitors; Triatoma; Trypanosoma cruzi; Animalia; Triatoma infestans; Trypanosoma cruzi
Año:2006
Volumen:132
Número:3
Página de inicio:345
Página de fin:355
DOI: http://dx.doi.org/10.1017/S0031182005009030
Título revista:Parasitology
Título revista abreviado:Parasitology
ISSN:00311820
CODEN:PARAA
CAS:concanavalin A, 11028-71-0; cysteine proteinase, 37353-41-6; n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine, 66701-25-5; tosyllysyl chloromethyl ketone, 2364-87-6; Culture Media; Cysteine Endopeptidases, EC 3.4.22.-; Protease Inhibitors
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00311820_v132_n3_p345_Duschak

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Citas:

---------- APA ----------
Duschak, V.G., Barboza, M., García, G.A., Lammel, E.M., Couto, A.S. & Isola, E.L.D. (2006) . Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis. Parasitology, 132(3), 345-355.
http://dx.doi.org/10.1017/S0031182005009030
---------- CHICAGO ----------
Duschak, V.G., Barboza, M., García, G.A., Lammel, E.M., Couto, A.S., Isola, E.L.D. "Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis" . Parasitology 132, no. 3 (2006) : 345-355.
http://dx.doi.org/10.1017/S0031182005009030
---------- MLA ----------
Duschak, V.G., Barboza, M., García, G.A., Lammel, E.M., Couto, A.S., Isola, E.L.D. "Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis" . Parasitology, vol. 132, no. 3, 2006, pp. 345-355.
http://dx.doi.org/10.1017/S0031182005009030
---------- VANCOUVER ----------
Duschak, V.G., Barboza, M., García, G.A., Lammel, E.M., Couto, A.S., Isola, E.L.D. Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis. Parasitology. 2006;132(3):345-355.
http://dx.doi.org/10.1017/S0031182005009030