Molecular and antigenic characterization of Trypanosoma cruzi TolT proteins.
Journal
PLoS neglected tropical diseases
ISSN: 1935-2735
Titre abrégé: PLoS Negl Trop Dis
Pays: United States
ID NLM: 101291488
Informations de publication
Date de publication:
03 2019
03 2019
Historique:
received:
16
10
2018
accepted:
14
02
2019
revised:
26
03
2019
pubmed:
15
3
2019
medline:
23
4
2019
entrez:
15
3
2019
Statut:
epublish
Résumé
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. 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. 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.
Sections du résumé
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.
Identifiants
pubmed: 30870417
doi: 10.1371/journal.pntd.0007245
pii: PNTD-D-18-01550
pmc: PMC6435186
doi:
Substances chimiques
Antigens, Protozoan
0
Membrane Proteins
0
Protozoan Proteins
0
TolT protein, Trypanosoma cruzi
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0007245Subventions
Organisme : NIAID NIH HHS
ID : R01 AI104531
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI123070
Pays : United States
Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
Références
Cell Signal. 1999 Jan;11(1):1-14
pubmed: 10206339
Int J Parasitol. 1999 May;29(5):749-57
pubmed: 10404271
Infect Immun. 1999 Sep;67(9):4603-12
pubmed: 10456906
Free Radic Biol Med. 2000 Feb 1;28(3):463-99
pubmed: 10699758
Int J Parasitol. 2001 May 1;31(5-6):472-81
pubmed: 11334932
Infect Immun. 2001 Dec;69(12):7946-9
pubmed: 11705983
Curr Pharm Des. 2002;8(4):269-85
pubmed: 11860366
Microbiol Mol Biol Rev. 2002 Mar;66(1):122-54; table of contents
pubmed: 11875130
Antioxid Redox Signal. 2003 Feb;5(1):133-8
pubmed: 12626125
Mol Biochem Parasitol. 2004 Jan;133(1):81-91
pubmed: 14668015
J Biol Chem. 2004 Apr 16;279(16):15860-9
pubmed: 14749325
Science. 2005 Jul 15;309(5733):404-9
pubmed: 16020724
Science. 2005 Jul 15;309(5733):409-15
pubmed: 16020725
Science. 2005 Jul 15;309(5733):416-22
pubmed: 16020726
Science. 2005 Jul 15;309(5733):436-42
pubmed: 16020728
Microbes Infect. 2006 Feb;8(2):401-9
pubmed: 16253534
Nat Rev Microbiol. 2006 Mar;4(3):229-36
pubmed: 16489349
J Mol Biol. 2007 Mar 9;366(5):1424-36
pubmed: 17222427
PLoS Pathog. 2008 Sep 05;4(9):e1000147
pubmed: 18773118
PLoS Negl Trop Dis. 2008 Oct 08;2(10):e316
pubmed: 18841200
Clin Chem. 2009 Apr;55(4):611-22
pubmed: 19246619
BMC Genomics. 2009 Aug 07;10:370
pubmed: 19664227
Nat Rev Microbiol. 2009 Nov;7(11):775-86
pubmed: 19806154
Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):5939-43
pubmed: 2068069
PLoS Negl Trop Dis. 2011 Mar 08;5(3):e984
pubmed: 21408126
Biochem J. 2011 Sep 1;438(2):303-13
pubmed: 21651499
Exp Parasitol. 1990 May;70(4):411-8
pubmed: 2182336
Mol Biochem Parasitol. 2012 Feb;181(2):61-72
pubmed: 22019385
BMC Evol Biol. 2011 Nov 17;11:334
pubmed: 22093578
J Proteome Res. 2012 Jan 1;11(1):237-46
pubmed: 22115061
Biochem J. 2012 Jun 1;444(2):211-8
pubmed: 22428617
J Biol Chem. 2012 Jul 27;287(31):26365-76
pubmed: 22707724
BMC Genomics. 2012 Oct 05;13:531
pubmed: 23035642
PLoS One. 2012;7(12):e50736
pubmed: 23284645
PLoS Negl Trop Dis. 2013 Nov 14;7(11):e2552
pubmed: 24244781
Genome Announc. 2014 Jan 30;2(1):null
pubmed: 24482508
Mol Cell Proteomics. 2014 Dec;13(12):3457-72
pubmed: 25225356
PLoS Negl Trop Dis. 2014 Sep 18;8(9):e3176
pubmed: 25233456
Cell Host Microbe. 2014 Oct 8;16(4):439-49
pubmed: 25299330
Microb Pathog. 2014 Dec;77:42-52
pubmed: 25448467
J Proteomics. 2015 Feb 6;115:58-65
pubmed: 25534883
Clin Vaccine Immunol. 2015 Mar;22(3):304-12
pubmed: 25589551
Semin Immunopathol. 2015 May;37(3):233-8
pubmed: 25921214
Mol Cell Proteomics. 2015 Jul;14(7):1871-84
pubmed: 25922409
PLoS One. 2015 Jun 18;10(6):e0130099
pubmed: 26086767
J Immunol Res. 2015;2015:178947
pubmed: 26240832
Biol Open. 2015 Aug 14;4(9):1143-53
pubmed: 26276100
J Extracell Vesicles. 2015 Nov 26;4:28734
pubmed: 26613751
Trends Parasitol. 2016 Apr;32(4):309-324
pubmed: 26776656
Biosens Bioelectron. 2016 Jun 15;80:24-33
pubmed: 26802749
PLoS Pathog. 2016 Apr 08;12(4):e1005559
pubmed: 27058585
PLoS Negl Trop Dis. 2016 Aug 29;10(8):e0004792
pubmed: 27571035
Trends Parasitol. 2017 Feb;33(2):102-112
pubmed: 27843019
Lancet Glob Health. 2017 Apr;5(4):e379-e380
pubmed: 28256341
Adv Parasitol. 2017;97:1-45
pubmed: 28325368
J Biol Chem. 2017 Aug 18;292(33):13584-13598
pubmed: 28642371
Lancet. 2018 Jan 6;391(10115):82-94
pubmed: 28673423
PLoS Negl Trop Dis. 2017 Aug 11;11(8):e0005856
pubmed: 28800609
PLoS Negl Trop Dis. 2017 Sep 29;11(9):e0005986
pubmed: 28961244
PLoS Negl Trop Dis. 2017 Oct 9;11(10):e0005972
pubmed: 28991925
Microb Genom. 2018 Feb 14;:null
pubmed: 29442617
J Extracell Vesicles. 2018 Apr 17;7(1):1463779
pubmed: 29696081
Microb Genom. 2018 May;4(5):
pubmed: 29708484
Sci Rep. 2018 Oct 2;8(1):14631
pubmed: 30279473
Mol Biochem Parasitol. 1998 Mar 1;91(1):37-49
pubmed: 9574924