Trypanosoma cruzi: Partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose

Vilma G. Duschak, Mariana Barboza Gardner, Alicia S. Couto

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)122-130
Number of pages9
JournalExperimental Parasitology
Volume104
Issue number3-4
DOIs
StatePublished - Jan 1 2003
Externally publishedYes

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Trypanosoma cruzi
Protein Isoforms
Tosyllysine Chloromethyl Ketone
Cystatins
Chromogenic Compounds
Aprotinin
Cysteine Proteases
Gelatin
Bovine Serum Albumin
Oligosaccharides
Affinity Chromatography
Lectins
Edetic Acid
Sepharose
Anions
cruzipain
concanavalin A-sepharose
Chromatography
Hemoglobins
Gels

ASJC Scopus subject areas

  • Parasitology
  • Immunology

Cite this

Trypanosoma cruzi : Partial characterization of minor cruzipain isoforms non-adsorbed to Concanavalin A-Sepharose. / Duschak, Vilma G.; Barboza Gardner, Mariana; Couto, Alicia S.

In: Experimental Parasitology, Vol. 104, No. 3-4, 01.01.2003, p. 122-130.

Research output: Contribution to journalArticle

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