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Title: Trypanosoma cruzi: isolation and characterization of aspartyl proteases
Authors: Pinho, Rosa T.
Beltramini, Leila M.
Alves, Carlos R.
De Simone, Salvatore Giovanni
Affilliation: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunologia Clínica. Rio de Janeiro, RJ, Brasil.
Universidade de São Paulo. Instituto de Física de São Carlos. Departamento de Física e Informática. São paulo, SP, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz., Laboratório de Bioquímica de Proteínas e Peptídeos. Rio de Janeiro, RJ, Brasil / Universidade Federal Fluminense. Instituto de Biologia. Departamento de Biologia Celular e Molecular. Niterói, RJ, Brasil.
Abstract: Two aspartyl proteases activities were identified and isolated from Trypanosoma cruzi epimastigotes: cruzipsin-I (CZP-I) and cruzipsin-II (CZP-II). One was isolated from a soluble fraction (CZP-II) and the other was solubilized with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CZP-I). The molecular mass of both proteases was estimated to be 120 kDa by HPLC gel filtration and the activity of the enzymes was detected in a doublet of bands (56 and 48 kDa) by substrate-sodium dodecyl sulphate-polyacrylamide-gelatin gel electrophoresis. Substrate specificity studies indicated that the enzymes consistently hydrolyze the cathepsin D substrate Phe-Ala-Ala-Phe (4-NO2)-Phe-Val-Leu-O4MP but failed to hydrolyze serine and other protease substrates. Both proteases activities were strongly inhibited by the classic inhibitor pepstatin-A (> or =68%) and the aspartic active site labeling agent, 1,2-epoxy-3-(phenyl-nitrophenoxy) propane (> or =80%). These findings show that both proteases are novel T. cruzi acidic proteases. The physiological function of these enzymes in T. cruzi has under investigation.
Keywords: Trypanosoma cruzi
Cathepsin D
keywords: Trypanosoma cruzi
Catepsina D
Issue Date: 2009
Publisher: Elsevier
Citation: PINHO, Rosa T. et al. Trypanosoma cruzi: Isolation and characterization of aspartyl proteases. Experimental Parasitology, v.122, n.2, p.128-133, 2009
DOI: 10.1016/j.exppara.2009.02.005
ISSN: 0014-4894
Copyright: restricted access
Appears in Collections:IOC - Artigos de Periódicos

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