Surface electrical charge of bloodstream trypomastigotes of Trypanosoma cruzi strains

Sousa, Maria Auxiliadora de

doi:10.1590/s0074-02761983000400014

Cited by 4 publications

(3 citation statements)

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“…Thereafter, the parasites were incubated in TAU3AAG medium (TAU supplemented with 10 mM L-proline, 50 mM Lglutamate, 2 mM L-aspartate, 10 mM glucose) to a final concentration 3 x 10 6 cell/ml in a final volume of 70 ml in Roux flask. They were incubated at 27°C for 72 hr, then centrifuged at 10,000 xg, resuspended in TAU medium, treated for 30 min at 37°C with fresh guinea pig serum, and separated on DEAE cellulose (Sousa 1983) as previously described (Contreras et al 1994). Metacyclic trypomastigotes from triatomine condition were obtained from epimastigote forms with no more than two months in culture.…”

Section: Methodsmentioning

confidence: 99%

Trypanosoma cruzi: Maintenance in Culture Modify Gene and Antigenic Expression of Metacyclic Trypomastigotes

Contreras

Lima²,

Zorrilla³

1998

Mem. Inst. Oswaldo Cruz

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Section: Methodsmentioning

confidence: 99%

Trypanosoma cruzi: Maintenance in Culture Modify Gene and Antigenic Expression of Metacyclic Trypomastigotes

Contreras

Lima²,

Zorrilla³

1998

Mem. Inst. Oswaldo Cruz

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“…Cell culture of T. cruzi trypomastigotes (Y strain) were maintained by infection of LLC-MK 2 cells, as described [19] in Minimum Essential Medium (MEM-GIBCO) containing 2% of Fetal Bovine Serum (FBS), under 5% CO 2 and 37˚C. Five days after infection of the host cell, culture supernatants containing the trypomastigotes were centrifuged and purified as described [20]. The commercially available extracellular matrix (Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix, Gibco, 150 μl ECM per sample) was used to incubate part of the trypomastigotes (5 x 10 8 cells in 5 ml MEM) as described [21].…”

Section: T Cruzi Trypomastigotes Cultivation and Incubation With Ecmmentioning

confidence: 99%

Global changes in nitration levels and DNA binding profile of Trypanosoma cruzi histones induced by incubation with host extracellular matrix

et al. 2020

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Adhesion of T. cruzi trypomastigotes to components of the extracellular matrix (ECM) is an important step in mammalian host cell invasion. We have recently described a significant increase in the tyrosine nitration levels of histones H2A and H4 when trypomastigotes are incubated with components of the ECM. In this work, we used chromatin immunoprecipitation (ChIP) with an anti-nitrotyrosine antibody followed by mass spectrometry to identify nitrated DNA binding proteins in T. cruzi and to detect alterations in nitration levels induced upon parasite incubation with the ECM. Histone H1, H2B, H2A and H3 were detected among the 9 most abundant nitrated DNA binding proteins using this proteomic approach. One nitrated tyrosine residue (Y29) was identified in Histone H2B in the MS/MS spectrum. In addition, we observed a significant increase in the nitration levels of histones H1, H2B, H2A and H4 upon parasite incubation with ECM. Finally, we used ChIP-Seq to map global changes in the DNA binding profile of nitrated proteins. We observed a significant change in the binding pattern of nitrated proteins to DNA after parasite incubation with ECM. This work provides the first global profile of nitrated DNA binding proteins in T. cruzi and additional evidence for modification in the nitration profile of histones upon parasite incubation with ECM. Our data also indicate that the parasite interaction with the ECM induces alterations in chromatin structure, possibly affecting nuclear functions.

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“…The epimastigote form can be readily obtained in axenic culture [11], and has been used for this reason in most biochemical studies performed on the parasite. Metacyclic trypomastigotes can be obtained by differentiation of epimastigotes in vitro [12], whereas amastigotes and trypomastigotes, resembling those found in the bloodstream of the mammal, can be obtained from infected mammalian cells [13]. Although the epimastigote form, and even non-pathogenic Trypanosomatids, such as Crithidia fasciculata, taken as models, have been used in the past, nowadays it is quite clear that any serious study of the effect of drugs on the parasite in vitro must include the stages present in the mammal.…”

Section: Introduction: Trypanosoma Cruzi and Chagas Diseasementioning

confidence: 99%

The Major Cysteine Proteinase of Trypanosoma cruzi: A Valid Target for Chemotherapy of Chagas Disease

Cazzulo

Stoka²,

Türk

2001

CPD

159

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Trypanosoma cruzi, the causative agent of the American Trypanosomiasis, Chagas disease, contains a major cysteine proteinase (CP), cruzipain (also known as cruzain, or GP57/51). The enzyme is a member of the papain C1 family of CPs, with a specificity intermediate between those of cathepsin L and cathepsin B. The enzyme, which is expressed at different levels by different parasite stages, is encoded by a high number of genes (up to 130 in the Tul2 strain), which code for a pre-pro-enzyme. Mature cruzipain consists of a catalytic moiety with high homology to cathepsins S and L, and a C-terminal domain, characteristic of Type I CPs of Trypanosomatids, and absent in all other C1 family CPs described so far. Irreversible inhibitors of cruzipain (peptidyl diazomethylketones, peptidyl fluoromethylketones, peptidyl vinyl sulphones) are able to block the differentiation steps in the parasite's life cycle, and effectively kill the organism. Recently, a vinyl sulphone derivative (N-piperazine-Phe-hPhe-vinyl sulphone phenyl) which is an efficient inhibitor of cruzipain and kills T. cruzi by inducing an accumulation of unprocessed cruzipain in the Golgi cisternae, interfering with the secretory pathway, has been tested in vivo in a mice model (J.H. McKerrow et al.). The curative effects observed, as well as the good bioavailability of the inhibitor and its apparent lack of undesirable side effects, make it a promising lead compound for the development of new drugs for the chemotherapy of Chagas disease.

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Surface electrical charge of bloodstream trypomastigotes of Trypanosoma cruzi strains

Cited by 4 publications

References 0 publications

Trypanosoma cruzi: Maintenance in Culture Modify Gene and Antigenic Expression of Metacyclic Trypomastigotes

Trypanosoma cruzi: Maintenance in Culture Modify Gene and Antigenic Expression of Metacyclic Trypomastigotes

Global changes in nitration levels and DNA binding profile of Trypanosoma cruzi histones induced by incubation with host extracellular matrix

The Major Cysteine Proteinase of Trypanosoma cruzi: A Valid Target for Chemotherapy of Chagas Disease

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