Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins.
BACKGROUND:TolT was originally described as a Trypanosoma cruzi molecule that accumulated on the trypomastigote flagellum bearing similarity to bacterial TolA colicins receptors. Preliminary biochemical studies indicated that TolT resolved in SDS-PAGE as ~3-5 different bands with sizes between 34 an...
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ftdoajarticles:oai:doaj.org/article:084386b17edb44459ea0f1c8f4f44583 2023-05-15T15:12:28+02:00 Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins. Maite Lobo Virginia Balouz Luciano Melli Giannina Carlevaro María E Cortina María de Los Milagros Cámara Gaspar E Cánepa Santiago J Carmona Jaime Altcheh Oscar Campetella Andrés E Ciocchini Fernán Agüero Juan Mucci Carlos A Buscaglia 2019-03-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0007245 https://doaj.org/article/084386b17edb44459ea0f1c8f4f44583 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC6435186?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0007245 https://doaj.org/article/084386b17edb44459ea0f1c8f4f44583 PLoS Neglected Tropical Diseases, Vol 13, Iss 3, p e0007245 (2019) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2019 ftdoajarticles https://doi.org/10.1371/journal.pntd.0007245 2022-12-31T15:19:02Z BACKGROUND:TolT was originally described as a Trypanosoma cruzi molecule that accumulated on the trypomastigote flagellum bearing similarity to bacterial TolA colicins receptors. Preliminary biochemical studies indicated that TolT resolved in SDS-PAGE as ~3-5 different bands with sizes between 34 and 45 kDa, and that this heterogeneity could be ascribed to differences in polypeptide glycosylation. However, the recurrent identification of TolT-deduced peptides, and variations thereof, in trypomastigote proteomic surveys suggested an intrinsic TolT complexity, and prompted us to undertake a thorough reassessment of this antigen. METHODS/PRINCIPLE FINDINGS:Genome mining exercises showed that TolT constitutes a larger-than-expected family of genes, with at least 12 polymorphic members in the T. cruzi CL Brener reference strain and homologs in different trypanosomes. According to structural features, TolT deduced proteins could be split into three robust groups, termed TolT-A, TolT-B, and TolT-C, all of them showing marginal sequence similarity to bacterial TolA proteins and canonical signatures of surface localization/membrane association, most of which were herein experimentally validated. Further biochemical and microscopy-based characterizations indicated that this grouping may have a functional correlate, as TolT-A, TolT-B and TolT-C molecules showed differences in their expression profile, sub-cellular distribution, post-translational modification(s) and antigenic structure. We finally used a recently developed fluorescence magnetic beads immunoassay to validate a recombinant protein spanning the central and mature region of a TolT-B deduced molecule for Chagas disease serodiagnosis. CONCLUSION/SIGNIFICANCE:This study unveiled an unexpected genetic and biochemical complexity within the TolT family, which could be exploited for the development of novel T. cruzi biomarkers with diagnostic/therapeutic applications. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 13 3 e0007245 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Maite Lobo Virginia Balouz Luciano Melli Giannina Carlevaro María E Cortina María de Los Milagros Cámara Gaspar E Cánepa Santiago J Carmona Jaime Altcheh Oscar Campetella Andrés E Ciocchini Fernán Agüero Juan Mucci Carlos A Buscaglia Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND:TolT was originally described as a Trypanosoma cruzi molecule that accumulated on the trypomastigote flagellum bearing similarity to bacterial TolA colicins receptors. Preliminary biochemical studies indicated that TolT resolved in SDS-PAGE as ~3-5 different bands with sizes between 34 and 45 kDa, and that this heterogeneity could be ascribed to differences in polypeptide glycosylation. However, the recurrent identification of TolT-deduced peptides, and variations thereof, in trypomastigote proteomic surveys suggested an intrinsic TolT complexity, and prompted us to undertake a thorough reassessment of this antigen. METHODS/PRINCIPLE FINDINGS:Genome mining exercises showed that TolT constitutes a larger-than-expected family of genes, with at least 12 polymorphic members in the T. cruzi CL Brener reference strain and homologs in different trypanosomes. According to structural features, TolT deduced proteins could be split into three robust groups, termed TolT-A, TolT-B, and TolT-C, all of them showing marginal sequence similarity to bacterial TolA proteins and canonical signatures of surface localization/membrane association, most of which were herein experimentally validated. Further biochemical and microscopy-based characterizations indicated that this grouping may have a functional correlate, as TolT-A, TolT-B and TolT-C molecules showed differences in their expression profile, sub-cellular distribution, post-translational modification(s) and antigenic structure. We finally used a recently developed fluorescence magnetic beads immunoassay to validate a recombinant protein spanning the central and mature region of a TolT-B deduced molecule for Chagas disease serodiagnosis. CONCLUSION/SIGNIFICANCE:This study unveiled an unexpected genetic and biochemical complexity within the TolT family, which could be exploited for the development of novel T. cruzi biomarkers with diagnostic/therapeutic applications. |
format |
Article in Journal/Newspaper |
author |
Maite Lobo Virginia Balouz Luciano Melli Giannina Carlevaro María E Cortina María de Los Milagros Cámara Gaspar E Cánepa Santiago J Carmona Jaime Altcheh Oscar Campetella Andrés E Ciocchini Fernán Agüero Juan Mucci Carlos A Buscaglia |
author_facet |
Maite Lobo Virginia Balouz Luciano Melli Giannina Carlevaro María E Cortina María de Los Milagros Cámara Gaspar E Cánepa Santiago J Carmona Jaime Altcheh Oscar Campetella Andrés E Ciocchini Fernán Agüero Juan Mucci Carlos A Buscaglia |
author_sort |
Maite Lobo |
title |
Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins. |
title_short |
Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins. |
title_full |
Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins. |
title_fullStr |
Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins. |
title_full_unstemmed |
Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins. |
title_sort |
molecular and antigenic characterization of trypanosoma cruzi tolt proteins. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2019 |
url |
https://doi.org/10.1371/journal.pntd.0007245 https://doaj.org/article/084386b17edb44459ea0f1c8f4f44583 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 13, Iss 3, p e0007245 (2019) |
op_relation |
http://europepmc.org/articles/PMC6435186?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0007245 https://doaj.org/article/084386b17edb44459ea0f1c8f4f44583 |
op_doi |
https://doi.org/10.1371/journal.pntd.0007245 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
13 |
container_issue |
3 |
container_start_page |
e0007245 |
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1766343139314892800 |