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Abstract:

The present paper reports the partial characterization of a subset of atypical cruzipain molecules which do not bind to Concanavalin A-Sepharose column. They are present in different strains of epimastigote forms of Trypanosoma cruzi and represent a 2-4% of total cruzipain. They were purified by affinity chromatography on Cystatin-Sepharose, recognized by the polyclonal anti-cruzipain serum, and their activity in gelatin-containing gels was completely abolished by E-64, TLCK, leupeptin, and aprotinin but not by PMSF, pepstatin A, EDTA or 1,10-phenantroline. These cysteine proteinases, as well as cruzipain showed to be endoproteinases able to hydrolize azocasein, hemoglobin, and bovine serum albumin at acidic pHs. However, evidences are presented indicating that this subset of cruzipain isoforms were also able to use the same blocked chromogenic peptidyl substrates than cruzipain at similar optimal alkaline pH values although with a different order of preference. Moreover, they showed a different oligosaccharide pattern after enzymatic treatment by high pH anion exchange chromatography, suggesting that this structural difference may account for the atypical behaviour in the lectin columm. © 2003 Elsevier Inc. All rights reserved.

Registro:

Documento: Artículo
Título:Trypanosoma cruzi: Partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose
Autor:Duschak, V.G.; Barboza, M.; Couto, A.S.
Filiación:Inst. de Invest. Biotecnologicas, Univ. Nac. de Gen. San Martín, INTI-Edificio 24, Av. Gral Paz y Albarellos, Prov. Buenos Aires, Argentina
CIHIDECAR, Depto. de Quimica Orgánica, University of Buenos Aires, Buenos Aires, Argentina
Inst. Nac. de Parasitología, Dr. Mario Fatala Chabén, Ministerio de Salud, Av. Paseo Colón 568 (1063), Buenos Aires, Argentina
Palabras clave:1,10 phenanthroline; aprotinin; azo compound; bovine serum albumin; casein; chromogenic substrate; concanavalin A; cruzipain; cystatin; cysteine proteinase; edetic acid; fluoride; gelatin; hemoglobin; isoenzyme; lectin; leupeptin; n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine; oligosaccharide; pepstatin; peptide derivative; phenylmethylsulfonylfluoride; polyclonal antibody; sepharose; tosyllysyl chloromethyl ketone; unclassified drug; acidity; adsorption; affinity chromatography; alkalinity; anion exchange chromatography; article; controlled study; enzyme activity; enzyme analysis; enzyme assay; enzyme purification; enzyme substrate; epimastigote; hydrolysis; nonhuman; priority journal; protein protein interaction; Trypanosoma cruzi; Bovinae; Trypanosoma; Trypanosoma cruzi
Año:2003
Volumen:104
Número:3-4
Página de inicio:122
Página de fin:130
DOI: http://dx.doi.org/10.1016/j.exppara.2003.08.001
Título revista:Experimental Parasitology
Título revista abreviado:Exp. Parasitol.
ISSN:00144894
CODEN:EXPAA
CAS:1,10 phenanthroline, 3829-86-5, 66-71-7; aprotinin, 11004-21-0, 12407-79-3, 50936-63-5, 52229-70-6, 58591-29-0, 9050-74-2, 9075-10-9, 9087-70-1; casein, 9000-71-9; concanavalin A, 11028-71-0; cruzipain; cystatin, 81989-95-9; cysteine proteinase, 37353-41-6; edetic acid, 150-43-6, 60-00-4; fluoride, 16984-48-8; gelatin, 9000-70-8; hemoglobin, 9008-02-0; leupeptin, 54577-99-0; n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine, 66701-25-5; pepstatin, 26305-03-3, 39324-30-6; sepharose, 61970-08-9; tosyllysyl chloromethyl ketone, 2364-87-6
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00144894_v104_n3-4_p122_Duschak

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Citas:

---------- APA ----------
Duschak, V.G., Barboza, M. & Couto, A.S. (2003) . Trypanosoma cruzi: Partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose. Experimental Parasitology, 104(3-4), 122-130.
http://dx.doi.org/10.1016/j.exppara.2003.08.001
---------- CHICAGO ----------
Duschak, V.G., Barboza, M., Couto, A.S. "Trypanosoma cruzi: Partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose" . Experimental Parasitology 104, no. 3-4 (2003) : 122-130.
http://dx.doi.org/10.1016/j.exppara.2003.08.001
---------- MLA ----------
Duschak, V.G., Barboza, M., Couto, A.S. "Trypanosoma cruzi: Partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose" . Experimental Parasitology, vol. 104, no. 3-4, 2003, pp. 122-130.
http://dx.doi.org/10.1016/j.exppara.2003.08.001
---------- VANCOUVER ----------
Duschak, V.G., Barboza, M., Couto, A.S. Trypanosoma cruzi: Partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose. Exp. Parasitol. 2003;104(3-4):122-130.
http://dx.doi.org/10.1016/j.exppara.2003.08.001