Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis

V. G. Duschak, Mariana Barboza Gardner, G. A. García, E. M. Lammel, A. S. Couto, E. L.D. Isola

Research output: Contribution to journalArticle

21 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)345-355
Number of pages11
JournalParasitology
Volume132
Issue number3
DOIs
StatePublished - Mar 1 2006
Externally publishedYes

Fingerprint

Cysteine Proteases
Trypanosoma cruzi
cysteine proteinases
cruzipain
Sepharose
agarose
Tosyllysine Chloromethyl Ketone
Peptide Hydrolases
Cystatins
Triatoma
Cysteine Proteinase Inhibitors
proteinases
Endopeptidases
epimastigotes
Triatoma infestans
trypomastigotes
Gelatin
cysteine proteinase inhibitors
cystatins
Oligosaccharides

Keywords

  • Cruzipain
  • Cysteine proteinases
  • Metacyclogenesis
  • Trypanosoma cruzi

ASJC Scopus subject areas

  • Parasitology
  • Animal Science and Zoology
  • Infectious Diseases

Cite this

Duschak, V. G., Barboza Gardner, 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. https://doi.org/10.1017/S0031182005009030

Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis. / Duschak, V. G.; Barboza Gardner, Mariana; García, G. A.; Lammel, E. M.; Couto, A. S.; Isola, E. L.D.

In: Parasitology, Vol. 132, No. 3, 01.03.2006, p. 345-355.

Research output: Contribution to journalArticle

Duschak, VG, Barboza Gardner, M, García, GA, Lammel, EM, Couto, AS & Isola, ELD 2006, 'Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis', Parasitology, vol. 132, no. 3, pp. 345-355. https://doi.org/10.1017/S0031182005009030
Duschak, V. G. ; Barboza Gardner, Mariana ; García, G. A. ; Lammel, E. M. ; Couto, A. S. ; Isola, E. L.D. / Novel cysteine proteinase in Trypanosoma cruzi metacyclogenesis. In: Parasitology. 2006 ; Vol. 132, No. 3. pp. 345-355.
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