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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Published in:PLOS Neglected Tropical Diseases
Main Authors: 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
Format: Article in Journal/Newspaper
Language:English
Published: Public Library of Science (PLoS) 2019
Subjects:
Online Access:https://doi.org/10.1371/journal.pntd.0007245
https://doaj.org/article/084386b17edb44459ea0f1c8f4f44583
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spelling 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
institution Open Polar
collection Directory of Open Access Journals: DOAJ Articles
op_collection_id ftdoajarticles
language English
topic Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
spellingShingle 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
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