Characterization and expression of proteases during Trypanosoma cruzi metacyclogenesis

Bonaldo, Myrna C.; d’Escoffier, Luiz Ney; Salles, Jussára M.; Goldenberg, Samuel

doi:10.1016/0014-4894(91)90006-i

Cited by 93 publications

(48 citation statements)

References 24 publications

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“…It was reported that cruzipain levels are significantly increased at early stages of parasite differentiation (Bonaldo et al, 1991) and that enzyme overexpression in epimastigotes (Silvio X10/6) increased metacyclogenesis (Tomás et al, 1997). Considering that the growth rate was not affected in chagasin-overexpressing cells, it is conceivable that parasite Smoothmuscle cells grown to semiconfluence were incubated in DMEM plus 2% FCS for 1 hour at 37°C, with buffer alone, buffer containing 10 µM E-64 or different concentrations of recombinant chagasin.…”

Section: Discussionmentioning

confidence: 99%

“…Cruzipain is posttranscriptionally regulated during the parasite's life cycle (Tomas and Kelly, 1996). T. cruzi replicative stages (epimastigotes and amastigotes) exhibit high contents of cruzipain, whereas the nondividing infective forms (trypomastigotes) express this enzyme at low levels (Bonaldo et al, 1991). As is true for other trypanosomatid CPs (Sanderson et al, 2000;Caffrey et al, 2001), the proteolytic excision of the N-terminal pro-domain of the zymogen precursor, pro-cruzipain, generates the enzymatically active protease (Eakin et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

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Chagasin, the endogenous cysteine-protease inhibitor ofTrypanosoma cruzi, modulates parasite differentiation and invasion of mammalian cells

Santos

Sant’Anna

Terres

et al. 2005

Journal of Cell Science

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Chagasin is a Trypanosoma cruzi protein that was recently characterized as a tight-binding inhibitor of papain-like cysteine proteases (CPs). Considering that parasite virulence and morphogenesis depend on the endogenous activity of lysosomal CPs of the cruzipain family, we sought to determine whether chagasin and cruzipain interact in the living cell. Ultrastructural studies showed that chagasin and cruzipain both localize to the Golgi complex and reservosomes (lysosome-like organelles), whereas free chagasin was found in small intracellular vesicles, suggesting that chagasin trafficking pathways might intersect with those of cruzipain. Taking advantage of the fact that sodium dodecyl sulphate and β-mercaptoethanol prevent binding between the isolated proteins but do not dismantle preformed cruzipain-chagasin complexes, we obtained direct evidence that chagasin-cruzipain complexes are indeed formed in epimastigotes. Chagasin transfectants (fourfold increase in CP inhibitory activity) displayed low rates of differentiation (metacyclogenesis) and exhibited increased resistance to a synthetic CP inhibitor. These phenotypic changes were accompanied by a drastic reduction of soluble cruzipain activity and by upregulated secretion of cruzipain-chagasin molecular complexes. Analysis of six T. cruzi strains revealed that expression levels of cruzipain and chagasin are variable, but the molar ratios are fairly stable (∼50:1) in most strains, with the exception of the G strain (5:1), which is poorly infective. On the same vein, we found that trypomastigotes overexpressing chagasin are less infective than wild-type parasites in vitro. The deficiency of chagasin overexpressers is caused by lower activity of membrane-associated CPs, because membranes recovered from wild-type trypomastigotes restored infectivity and this effect was nullified by the CP inhibitor E-64. In summary, our studies suggest that chagasin regulates the endogenous activity of CP, thus indirectly modulating proteolytic functions that are essential for parasite differentiation and invasion of mammalian cells.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Chagasin, the endogenous cysteine-protease inhibitor ofTrypanosoma cruzi, modulates parasite differentiation and invasion of mammalian cells

Santos

Sant’Anna

Terres

et al. 2005

Journal of Cell Science

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“…Trypanosorna cruzi, a human pathogen in Central and South America, contains a metallo-protease which is mainly located in the cell membrane (Greig & Ashall 1990, Bonaldo et al 1991. T cruzi trypomastigotes penetrate host cells and a surface or secreted metalloprotease is presumed to be involved in penetration of cells (Piras et al 1985, Greig & Ashall 1990, Bonaldo et al 1991. The metallo-protease in trophozoite of Entarnoeba histolytica, a human pathogen, readily degrades host collagens, suggesting that it also plays an important role in the invasion of host tissue (Munoz et al 1982(Munoz et al , 1990.…”

Section: Discussionmentioning

confidence: 99%

Purified metallo-protease from the pathogenic haemoflagellate Cryptobia salmositica and its in vitro proteolytic activities

Zuo¹,

Woo²

1997

Dis. Aquat. Org.

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The metallo-protease (200 kDa) from Cryptobia salmositica was punfied using ionexchange chromatography and gelfiltration. The purity of the enzyme was confirmed as there was only one homogeneous band using SDS-PAGE. Under in vitro conditions the purified metallo-protease completely degraded the extracellular matrix proteins (collagen types I. [V. V and laminin) and the membrane proteins isolated from rainbow trout erythrocytes. The results confirm that the C. salmositica metallo-protease is a histolytic enzyme and it contributes to salmonid cryptobiosis and to the direct transmission of the parasite between fish.

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“…Metallopeptidases homologous to the gp63 of Leishmania spp. are present in T. cruzi, as well as two metallocarboxypeptidases belonging to the M32 family, before found only in prokaryotes (Bonaldo, 1991;Lowndes, 1996;Burleigh, 2002;Cuevas, 2003;Nogueira de Melo, 2004;Gutierrez, 2008;Niemirowicz, 2007;Niemirowicz, 2008;Kulkarni, 2009). The most abundant among these enzymes is cruzipain (Cz), also known as GP57/51 or cruzain (the recombinant catalytic domain of cruzipain) a cysteine peptidase (CP) expressed as a mixture of isoforms.…”

Section: Peptidasesmentioning

confidence: 99%

Novel Therapeutic Strategies for Chagas` Disease

Vermelho¹,

Fraga²,

Carvalho³

et al. 2011

Drug Development - A Case Study Based Insight Into Modern Strategies

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Characterization and expression of proteases during Trypanosoma cruzi metacyclogenesis

Cited by 93 publications

References 24 publications

Chagasin, the endogenous cysteine-protease inhibitor ofTrypanosoma cruzi, modulates parasite differentiation and invasion of mammalian cells

Chagasin, the endogenous cysteine-protease inhibitor ofTrypanosoma cruzi, modulates parasite differentiation and invasion of mammalian cells

Purified metallo-protease from the pathogenic haemoflagellate Cryptobia salmositica and its in vitro proteolytic activities

Novel Therapeutic Strategies for Chagas` Disease

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