Identification of O-Glcnacylated Proteins in Trypanosoma cruzi.

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It is caused by the protozoan Trypanosoma cruzi. Differences in disease severity have been attributed to a natural pleomorphism in T. cruzi. Several post-translational modifications (PTMs) have been studied in T. cruzi, but to date no work has focused on O-GlcNAcylation, a highly conserved monosaccharide-PTM of serine and threonine residues mainly found in nucleus, cytoplasm, and mitochondrion proteins. O-GlcNAcylation is thought to regulate protein function analogously to protein phosphorylation; indeed, crosstalk between both PTMs allows the cell to regulate its functions in response to nutrient levels and stress. Herein, we demonstrate O-GlcNAcylation in T. cruzi epimastigotes by three methods: by using specific antibodies against the modification in lysates and whole parasites, by click chemistry labeling, and by proteomics. In total, 1,271 putative O-GlcNAcylated proteins and six modification sequences were identified by mass spectrometry (data available via ProteomeXchange, ID PXD010285). Most of these proteins have structural and metabolic functions that are essential for parasite survival and evolution. Furthermore, O-GlcNAcylation pattern variations were observed by antibody detection under glucose deprivation and heat stress conditions, supporting their possible role in the adaptive response. Given the numerous biological processes in which O-GlcNAcylated proteins participate, its identification in T. cruzi proteins opens a new research field in the biology of Trypanosomatids, improve our understanding of infection processes and may allow us to identify new therapeutic targets.", "claims": [{"id": "claim-method", "statement": "False discovery rate (FDR) was processed using Percolator a semi-supervised machine learning to discriminate correct from incorrect peptide-spectrum matches (22), and was set to a q-value of 1 and 5% for, respectively, define high and low confident peptides.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:30984116", "locator": "sections[13] \"Protein Identification and Abundance Quantification\" paragraph 2", "quote": "False discovery rate (FDR) was processed using Percolator a semi-supervised machine learning to discriminate correct from incorrect peptide-spectrum matches (22), and was set to a q-value of 1 and 5% for, respectively, define high and low confident peptides."}], "state": "complete"}, {"id": "claim-data-access", "statement": "Mass spectrometry proteomics data were deposited and can be found in the ProteomeXchange Consortium via the PRIDE (25) partner repository with the dataset identifier PXD010285.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:30984116", "locator": "sections[20] \"Data Availability\" paragraph 2", "quote": "Mass spectrometry proteomics data were deposited and can be found in the ProteomeXchange Consortium via the PRIDE (25) partner repository with the dataset identifier PXD010285."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": null, "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:30984116"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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