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Proteome-wide modulation of S-nitrosylation in Trypanosoma cruzi trypomastigotes upon interaction with the host extracellular matrix

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Autor(es):
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]
Número total de Autores: 9
Afiliação do(s) autor(es):
[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
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: JOURNAL OF PROTEOMICS; v. 231, JAN 16 2021.
Citações Web of Science: 0
Resumo

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)

Processo FAPESP: 14/25494-9 - Resposta de Trypanosoma cruzi ao meio ambiente: matriz extracelular e mudanças de pH
Beneficiário:Maria Julia Manso Alves
Modalidade de apoio: Auxílio à Pesquisa - Regular
Processo FAPESP: 14/10046-0 - O envolvimento das modificações NO-dependentes no controle da transcrição gênica de Trypanosoma cruzi durante sua adesão à matriz extracelular do hospedeiro
Beneficiário:Rubens Daniel Miserani Magalhães
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 18/03727-2 - Papel dos segundos mensageiros nas vias de sinalização envolvidas na interação de Trypanosoma cruzi e matriz extracelular
Beneficiário:Nubia Carolina Manchola Varón
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 17/04032-5 - Dissecando a patogênese da Doença de Chagas através de abordagens glicômicas e glicoproteômicas
Beneficiário:Simon Ngao Mule
Modalidade de apoio: Bolsas no Brasil - Doutorado
Processo FAPESP: 18/18257-1 - EMU concedido no processo 14/06863-3: sistema de cromatografia líquida configurado para análise de carboidratos, aminoácidos, peptídeos e glicoproteínas
Beneficiário:Giuseppe Palmisano
Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
Processo FAPESP: 18/13283-4 - Descoberta de potências biomarcadores da Doença de Chagas em urina utilizando técnicas de espectrometria de massas
Beneficiário:Gilberto Santos de Oliveira
Modalidade de apoio: Bolsas no Brasil - Doutorado
Processo FAPESP: 18/15549-1 - Modificações pós-traducionais nos processos biológicos e no diagnóstico da Doença de Chagas: novas abordagens metodológicas e implicações biológicas
Beneficiário:Giuseppe Palmisano
Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores - Fase 2
Processo FAPESP: 12/03887-3 - Identificação de proteínas de Trypanosoma cruzi modificadas por S-nitrosilação e nitração após adesão com matriz extracelular.
Beneficiário:Milton César de Almeida Pereira
Modalidade de apoio: Bolsas no Brasil - Mestrado