| Full text | |
| Author(s): |
Mule, S. N.
[1]
;
Manchola, N. C.
[2]
;
de Oliveira, G. S.
[1]
;
Pereira, M.
[2, 3]
;
Magalhaes, R. D. M.
[4, 2]
;
Teixeira, A. A.
[5, 2]
;
Colli, W.
[2]
;
Alves, M. J. M.
[2]
;
Palmisano, G.
[1]
Total Authors: 9
|
| Affiliation: | [1] Univ Sao Paulo, Inst Ciencias Biomed, Dept Parasitol, Sao Paulo - Brazil
[2] Univ Sao Paulo, Dept Bioquim, Inst Quim, 748 Butanta, BR-05508000 Sao Paulo, SP - Brazil
[3] Univ Massachusetts, Sch Med, Div Infect Dis & Immunol, Worcester, MA 01605 - USA
[4] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Biol Celular & Mol, Ribeirao Preto, SP - Brazil
[5] New Mexico Consortium, Los Alamos, NM - USA
Total Affiliations: 5
|
| Document type: | Journal article |
| Source: | JOURNAL OF PROTEOMICS; v. 231, JAN 16 2021. |
| Web of Science Citations: | 0 |
| Abstract | |
Trypanosoma cruzi trypomastigotes adhere to extracellular matrix (ECM) to invade mammalian host cells regulating intracellular signaling pathways. Herein, resin-assisted enrichment of thiols combined with mass spectrometry were employed to map site-specific S-nitrosylated (SNO) proteins from T. cruzi trypomastigotes incubated (MTy) or not (Ty) with ECM. We confirmed the reduction of S-nitrosylation upon incubation with ECM, associated with a rewiring of the subcellular distribution and intracellular signaling pathways. Forty, 248 and 85 SNO-peptides were identified only in MTy, Ty or in both conditions, respectively. SNO proteins were enriched in ribosome, transport, carbohydrate and lipid metabolisms. Nitrosylation of histones H2B and H3 on Cys64 and Cys126, respectively, is described. Protein-protein interaction networks revealed ribosomal proteins, proteins involved in carbon and fatty acid metabolism to be among the enriched protein complexes. Kinases, phosphatases and enzymes involved in the metabolism of carbohydrates, lipids and amino acids were identified as nitrosylated and phosphorylated, suggesting a post-translational modifications crosstalk. In silico mapping of nitric oxide synthase (NOS) genes, previously uncharacterized, matched to four putative T. cruzi proteins expressing C-terminal NOS domain. Our results provide the first site-specific characterization of S-nitrosylated proteins in T. cruzi and their modulation upon ECM incubation before infection of the mammalian hosts. Significance: Protein S-nitrosylation represents a major molecular mechanism for signal transduction by nitric oxide. We present for the first time a proteomic profile of S-nitrosylated proteins from infective forms of T. cruzi, showing a decrease in SNO proteins after incubation of the parasite with the extracellular matrix, a necessary step for the parasite invasion of the host mammalian cells. We also show for the first time nitrosylation of H2B (Cys64) and H3 (Cys126) histones, sites not conserved in higher eukaryotic cells, and suggest that some specific histone isoforms are sensitive to NO signaling. S-nitrosylation in H2B and H3 histones are more abundant in MTy. Moreover, proteins involved in translation, glycolytic pathway and fatty acid metabolism are enriched in the present dataset. Comparison of the SNO proteome and the phosphoproteome, obtained previously under the same experimental conditions, show that most of the proteins sharing both modifications are involved in metabolic pathways, transport and ribosome function. The data suggest that both PTMs are involved in reprogramming the metabolism of T. cruzi in response to environmental changes. Although NO synthesis was detected in T. cruzi, the identification of NOS remains elusive. Analysis in silico showed two genes similar in domains to NADPH-dependent cytochrome-P450 reductase and two putative oxidoreductases, but no oxygenase domain of NOS was mapped in the T. cruzi genome. It is tempting to speculate that NO synthase-like from T. cruzi and its early NO-mediated pathways triggered in response to host interaction constitute potential diagnostic and therapeutic targets. (AU) | |
| FAPESP's process: | 14/25494-9 - Trypanosoma Cruzi signaling due to changes in the external conditions: ECM and pH changes |
| Grantee: | Maria Julia Manso Alves |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 14/10046-0 - Involvement of NO-dependent modifications in the control of Trypanosoma cruzi gene transcription throughout the adhesion of the parasite to the host extracellular matrix |
| Grantee: | Rubens Daniel Miserani Magalhães |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 18/03727-2 - The role of second messengers in the signaling pathways of the Trypanosoma Cruzi interaction with the extracellular matrix |
| Grantee: | Nubia Carolina Manchola Varón |
| Support Opportunities: | Scholarships in Brazil - Post-Doctoral |
| FAPESP's process: | 17/04032-5 - Dissecting the pathogenesis of Chagas Disease by deep glycomics and glycoproteomics approaches |
| Grantee: | Simon Ngao Mule |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 18/18257-1 - Multi-user equipment approved in grant 14/06863-3: HPLC system configured for analysis of carbohydrates, amino acidis, peptides and glycoproteins |
| Grantee: | Giuseppe Palmisano |
| Support Opportunities: | Multi-user Equipment Program |
| FAPESP's process: | 18/13283-4 - Discovery of biomarkers of Chagas' disease in urine using mass spectrometry techniques |
| Grantee: | Gilberto Santos de Oliveira |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 18/15549-1 - Post-translational modifications in Chagas Disease biological processes and diagnostics: novel methodological approaches and biological applications |
| Grantee: | Giuseppe Palmisano |
| Support Opportunities: | Research Grants - Young Investigators Grants - Phase 2 |
| FAPESP's process: | 12/03887-3 - Identification of Trypanosoma cruzi proteins modified by S-nitrosylation and nitration after adhesion to extracellular matrix. |
| Grantee: | Milton César de Almeida Pereira |
| Support Opportunities: | Scholarships in Brazil - Master |