Trypanosoma cruzi Presenilin-Like Transmembrane Aspartyl Protease: Characterization and Cellular Localization

Lechuga, Guilherme Curty; Napoleão-Pêgo, Paloma; Bottino, Carolina C.G.; Pinho, Rosa Teixeira de; Provance, David William; De-Simone, Salvatore Giovanni

doi:10.3390/biom10111564

Cited by 14 publications

(17 citation statements)

References 76 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…together with its fractionation to membranes, suggest that the NICT-like protein could also be localized to the endoplasmic reticulum, Golgi complex and mitochondria similar to the T. cruzi presenilin localization [14]. In other cellular systems, the integration of these organelles with PS/γ-secretase complex is responsible for aspects of the secretory pathways [36,37].…”

Section: Discussionmentioning

confidence: 82%

“…As epimastigotes are replicative forms, it would be expected that they would have a larger demand for molecules to sustain proliferation. Recently, we have showed that the replicative forms of T. cruzi has the highest levels of PS-like signals, and these results were associated with a potential role for a γ-secretase complex in functions related to proliferation and protein maturation [14].…”

Section: Discussionmentioning

confidence: 95%

“…While much is known about the GSC compounds in superior eukaryotic organisms, very little is known about its role in inferior eukaryotic cells. Recently, we have shown the potential for the presence of a GSC in Trypanosoma cruzi based on our characterization of a TM aspartyl protease that shows hallmarks of human PS [14]. T. cruzi is the causative agent for American trypanosomiasis, more commonly referred to as Chagas disease, and infections are typically spread by triatomine vectors that affects millions of people throughout Latin America [15].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Nicastrin-like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

Lechuga¹,

Napoleão-Pêgo²,

Gomes³

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Nicastrin (NICT) is a transmembrane protein physically associated with the polytypical aspartyl protease presenilin that plays a vital role in the correct localization and stabilization of presenilin to the membrane-bound γ-secretase complex. This complex is involved in the regulation of a wide range of cellular events including cell signaling and the regulation of endocytosed membrane proteins for their trafficking and protein processing. Mehtods: In Trypanosoma cruzi, the causal agent of the Chagas disease, an NICT-like protein (Tc/NICT) was identified with a short C-terminus orthologous to the human protein, a large ectodomain (ECD) with numerous glycosylation sites and a single core transmembrane domain containing a putative TM-domain (457GSVGA461) important for the γ-secretase complex activity. Results: Using the Spot-synthesis strategy with Chagasic patient sera, five extracellular epitopes were identified and synthetic forms were used to generate rabbit anti-Tc/NICT polyclonal serum that recognized a ~72-kDa molecule in immunoblots of T. cruzi epimastigote extracts. Confocal microscopy suggests that Tc/NICT is localized in the flagellar pocket, which is consistent with data from our previous studies with a T. cruzi presenilin-like protein. Phylogenetically, Tc/NICT was localized within a subgroup with the T. rangeli protein that are clearly detached from the other Trypanosomatidae such as T. brucei. These results, together with a comparative analysis of the selected peptide sequence regions between the T. cruzi and mammalian proteins suggest a divergence from the human NICT that might be relevant to Chagas disease pathology. As a whole, our data show that an NICT-like protein is expressed in the infective and replicative stages of T. cruzi and may be considered further evidence for a γ-secretase complex in trypanosomatids.

show abstract

Section: Discussionmentioning

confidence: 82%

Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Nicastrin-like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

Lechuga¹,

Napoleão-Pêgo²,

Gomes³

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…To verify the diagnostic performance of the peptides, sera from ninety-two DTP vaccinated individuals were tested on an ELISA-peptide based assay ( Figure 3 A,B). All 92 DTP-positive human sera and 38 mice sera reacted with the highly conserved DTx2–12 and DTx4–13 peptides but did not react with peptide EP7 [ 56 ] from T. cruzi (PPGEDMHTRDGPRE), considered by a scale value as a good test. Both CB/DTx2–12 and CB/DTx4–13 antigens demonstrated 100 and 99.96% sensitivity, respectively, and 100% specificity ( Figure 3 C,D).…”

Section: Resultsmentioning

confidence: 99%

Epitope Mapping of the Diphtheria Toxin and Development of an ELISA-Specific Diagnostic Assay

et al. 2021

Self Cite

View full text Add to dashboard Cite

Background: The diphtheria toxoid antigen is a major component in pediatric and booster combination vaccines and is known to raise a protective humoral immune response upon vaccination. Although antibodies are considered critical for diphtheria protection, little is known about the antigenic determinants that maintain humoral immunity. Methods: One-hundred and twelve 15 mer peptides covering the entire sequence of diphtheria toxin (DTx) protein were prepared by SPOT synthesis. The immunoreactivity of membrane-bound peptides with sera from mice immunized with a triple DTP vaccine allowed mapping of continuous B-cell epitopes, topological studies, multiantigen peptide (MAP) synthesis, and Enzyme-Linked Immunosorbent Assay (ELISA) development. Results: Twenty epitopes were identified, with two being in the signal peptide, five in the catalytic domain (CD), seven in the HBFT domain, and five in the receptor-binding domain (RBD). Two 17 mer (CB/Tx-2/12 and CB/DTx-4–13) derived biepitope peptides linked by a Gly-Gly spacer were chemically synthesized. The peptides were used as antigens to coat ELISA plates and assayed with human (huVS) and mice vaccinated sera (miVS) for in vitro diagnosis of diphtheria. The assay proved to be highly sensitive (99.96%) and specific (100%) for huVS and miVS and, when compared with a commercial ELISA test, demonstrated a high performance. Conclusions: Our work displayed the complete picture of the linear B cell IgG response epitope of the DTx responsible for the protective effect and demonstrated sufficient specificity and eligibility for phase IIB studies of some epitopes to develop new and fast diagnostic assays.

show abstract

“…While much is known about the GSC compounds in superior eukaryotic organisms, very little is known about their role in inferior eukaryotic cells. Recently, we showed the potential for the presence of a GSC in Trypanosoma cruzi based on our characterization of a TM aspartyl protease that shows hallmarks of human PS [14]. T. cruzi is the causative agent for American trypanosomiasis, more commonly referred to as Chagas disease, and infections are typically spread by triatomine vectors that affect millions of people throughout Latin America [15].…”

Section: Introductionmentioning

confidence: 99%

Nicastrin-Like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

et al. 2021

Self Cite

View full text Add to dashboard Cite

Nicastrin (NICT) is a transmembrane protein physically associated with the polytypical aspartyl protease presenilin that plays a vital role in the correct localization and stabilization of presenilin to the membrane-bound γ-secretase complex. This complex is involved in the regulation of a wide range of cellular events, including cell signaling and the regulation of endocytosed membrane proteins for their trafficking and protein processing. Methods: In Trypanosoma cruzi, the causal agent of the Chagas disease, a NICT-like protein (Tc/NICT) was identified with a short C-terminus orthologous to the human protein, a large ectodomain (ECD) with numerous glycosylation sites and a single-core transmembrane domain containing a putative TM-domain (457GSVGA461) important for the γ-secretase complex activity. Results: Using the Spot-synthesis strategy with Chagasic patient sera, five extracellular epitopes were identified and synthetic forms were used to generate rabbit anti-Tc/NICT polyclonal serum that recognized a ~72-kDa molecule in immunoblots of T. cruzi epimastigote extracts. Confocal microscopy suggests that Tc/NICT is localized in the flagellar pocket, which is consistent with data from our previous studies with a T. cruzi presenilin-like protein. Phylogenetically, Tc/NICT was localized within a subgroup with the T. rangeli protein that is clearly detached from the other Trypanosomatidae, such as T. brucei. These results, together with a comparative analysis of the selected peptide sequence regions between the T. cruzi and mammalian proteins, suggest a divergence from the human NICT that might be relevant to Chagas disease pathology. As a whole, our data show that a NICT-like protein is expressed in the infective and replicative stages of T. cruzi and may be considered further evidence for a γ-secretase complex in trypanosomatids.

show abstract

Trypanosoma cruzi Presenilin-Like Transmembrane Aspartyl Protease: Characterization and Cellular Localization

Cited by 14 publications

References 76 publications

Nicastrin-like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

Nicastrin-like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

Epitope Mapping of the Diphtheria Toxin and Development of an ELISA-Specific Diagnostic Assay

Nicastrin-Like, a Novel Transmembrane Protein from Trypanosoma cruzi Associated to the Flagellar Pocket

Contact Info

Product

Resources

About