Reevaluating the Trypanosoma cruzi proteomic map: The shotgun description of bloodstream trypomastigotes

Brunoro, Giselle Villa Flor; Caminha, Marcelle Almeida; Ferreira, André Teixeira da Silva; Leprevost, Felipe da Veiga; Carvalho, Paulo C.; Perales, Jonás; Valente, Richard H.; Menna-Barreto, Rubem Figueiredo Sadok

doi:10.1016/j.jprot.2014.12.003

Cited by 43 publications

(43 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…lack mobile retrotransposon elements (69). A recent proteomic analysis identified ϳ39 RHS isoforms that were expressed by T. cruzi bloodstream trypomastigotes (70); however, the diversity of RHS proteins detected in EV preparations was much more restricted, as only 8 full-length RHS variants were observed (Table S3). In ELISAs, only the recombinant RHS showed a strong response with immune sera from patients with various clinical degrees of Chagas disease.…”

Section: Discussionmentioning

confidence: 99%

Characterization and Diagnostic Application of Trypanosoma cruzi Trypomastigote Excreted-Secreted Antigens Shed in Extracellular Vesicles Released from Infected Mammalian Cells

et al. 2017

View full text Add to dashboard Cite

Chagas disease, caused by Trypanosoma cruzi, although endemic in many parts of Central and South America, is emerging as a global health threat through the potential contamination of blood supplies. Consequently, in the absence of a gold standard assay for the diagnosis of Chagas disease, additional antigens or strategies are needed. A proteomic analysis of the trypomastigote excreted-secreted antigens (TESA) associated with exosomal vesicles shed by T. cruzi identified ϳ80 parasite proteins, with the majority being trans-sialidases. Mass spectrometry analysis of immunoprecipitation products performed using Chagas immune sera showed a marked enrichment in a subset of TESA proteins. Of particular relevance for diagnostic applications were the retrotransposon hot spot (RHS) proteins, which are absent in Leishmania spp., parasites that often confound diagnosis of Chagas disease. Interestingly, serological screens using recombinant RHS showed a robust immunoreactivity with sera from patients with clinical stages of Chagas ranging from asymptomatic to advance cardiomyopathy and this immunoreactivity was comparable to that of crude TESA. More importantly, no cross-reactivity with RHS was detected with sera from patients with malaria, leishmaniasis, toxoplasmosis, or African sleeping sickness, making this protein an attractive reagent for diagnosis of Chagas disease.

show abstract

Section: Discussionmentioning

confidence: 99%

Characterization and Diagnostic Application of Trypanosoma cruzi Trypomastigote Excreted-Secreted Antigens Shed in Extracellular Vesicles Released from Infected Mammalian Cells

et al. 2017

View full text Add to dashboard Cite

show abstract

“…In our recent description of the proteomic map of bloodstream trypomastigotes, we discussed the presence of blood components, especially the plasma, erythrocytes and platelets of Mus musculus in these samples [27]. Even the high stringency of the parasite purification process is not enough to avoid the identification of host proteins.…”

Section: Discussionmentioning

confidence: 99%

“…cruzi bloodstream trypomastigotes (the Y strain) were obtained by performing a heart puncture on infected albino Swiss mice ( Mus musculus ) at peak parasitemia (7 th day) as previously described [27]. Six independent infections and heart punctures were performed in different days to generate six distinct pool of parasites which were considered distinct biological replicates.…”

Section: Methodsmentioning

confidence: 99%

“…T . cruzi protein entries (5,901 of them) were described in almost all the cellular compartments and metabolic pathways of the parasite, with 2,202 protein entries exclusively detected in the bloodstream forms in comparison with the culture-derived and metacyclic trypomastigote proteomic profiles reported in the literature [27], which justifies the use of bloodstream trypomastigotes as a model for pathogenesis and chemotherapy studies. In the present work, we further evaluated the mechanisms of action of the naphthoimidazoles in bloodstream trypomastigotes, which are the clinically relevant forms of the parasite, by differential gel electrophoresis (DIGE).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Differential Gel Electrophoresis (DIGE) Evaluation of Naphthoimidazoles Mode of Action: A Study in Trypanosoma cruzi Bloodstream Trypomastigotes

et al. 2016

Self Cite

View full text Add to dashboard Cite

BackgroundThe obligate intracellular protozoan Trypanosoma cruzi is the causative agent of Chagas disease, a neglected illness affecting millions of people in Latin America that recently entered non-endemic countries through immigration, as a consequence of globalization. The chemotherapy for this disease is based mainly on benznidazole and nifurtimox, which are very efficient nitroderivatives against the acute stage but present limited efficacy during the chronic phase. Our group has been studying the trypanocidal effects of naturally occurring quinones and their derivatives, and naphthoimidazoles derived from β-lapachone N1, N2 and N3 were the most active. To assess the molecular mechanisms of action of these compounds, we applied proteomic techniques to analyze treated bloodstream trypomastigotes, which are the clinically relevant stage of the parasite.Methodology/Principal FindingsThe approach consisted of quantification by 2D-DIGE followed by MALDI-TOF/TOF protein identification. A total of 61 differentially abundant protein spots were detected when comparing the control with each N1, N2 or N3 treatment, for 34 identified spots. Among the differentially abundant proteins were activated protein kinase C receptor, tubulin isoforms, asparagine synthetase, arginine kinase, elongation factor 2, enolase, guanine deaminase, heat shock proteins, hypothetical proteins, paraflagellar rod components, RAB GDP dissociation inhibitor, succinyl-CoA ligase, ATP synthase subunit B and methionine sulfoxide reductase.Conclusion/SignificanceOur results point to different modes of action for N1, N2 and N3, which indicate a great variety of metabolic pathways involved and allow for novel perspectives on the development of trypanocidal agents.

show abstract

“…Similarly, Trypanosoma cruzi and Leishmania species alternate between hosts with dividing and non-dividing stages, but are intracellular in the mammal host and extracellular in the insect. To accommodate these widely differing environments with respect to temperature, nutrients and immune interactions numerous differences in protein abundance between developmental stages have been identified [31–38]. Indeed, proteomic analyses suggest that as many as one-third of all proteins show at least a two-fold difference in abundance in T. brucei cultured BF versus cultured PF (PCF) [31,33].…”

Section: Trypanosomatids: Managing a Complex Life Cycle Without Transmentioning

confidence: 99%

Illuminating Parasite Protein Production by Ribosome Profiling

Parsons

Myler

2016

Trends in Parasitology

View full text Add to dashboard Cite

While technologies for global enumeration of transcript abundance are well-developed, those that assess protein abundance require tailoring to penetrate to low abundance proteins. Ribosome profiling circumvents this challenge by measuring global protein production via sequencing small mRNA fragments protected by the assembled ribosome. This powerful approach is now being applied to protozoan parasites, including trypanosomes and Plasmodium. It has been used to identify new protein coding sequences (CDSs) and clarify the boundaries of previously annotated CDSs in Trypanosoma brucei. Ribosome profiling has demonstrated that translation efficiencies vary widely between genes and, for trypanosomes at least, for the same gene across stages. The ribosomal proteins are themselves subjected to translational control, suggesting a means of reinforcing global translational regulation.

show abstract

Reevaluating the Trypanosoma cruzi proteomic map: The shotgun description of bloodstream trypomastigotes

Cited by 43 publications

References 47 publications

Characterization and Diagnostic Application of Trypanosoma cruzi Trypomastigote Excreted-Secreted Antigens Shed in Extracellular Vesicles Released from Infected Mammalian Cells

Characterization and Diagnostic Application of Trypanosoma cruzi Trypomastigote Excreted-Secreted Antigens Shed in Extracellular Vesicles Released from Infected Mammalian Cells

Differential Gel Electrophoresis (DIGE) Evaluation of Naphthoimidazoles Mode of Action: A Study in Trypanosoma cruzi Bloodstream Trypomastigotes

Illuminating Parasite Protein Production by Ribosome Profiling

Contact Info

Product

Resources

About