Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:24267406", "kind": "science-record", "revision": 7, "content_hash": "sha256:097aca30fa4532df9ff0764840395c70bfbddf594a2e1aa37d80f389c1ca2c9c", "created_at": "2026-11-01T15:40:03Z", "updated_at": "2026-11-01T21:58:14Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics.", "doi": "10.1186/1756-3305-6-335", "pmid": "24267406", "authors": ["Letícia Pollo-Oliveira", "Harm Post", "Marcio Luis Acencio", "Ney Lemke", "Henk van den Toorn", "Vinicius Tragante", "Albert J R Heck", "A F Maarten Altelaar", "Ana Patrícia Yatsuda"], "journal": "Parasites & vectors", "year": 2013, "abstract": "BACKGROUND: The apicomplexan parasite Neospora caninum causes neosporosis, a disease that leads to abortion or stillbirth in cattle, generating an economic impact on the dairy and beef cattle trade. As an obligatory intracellular parasite, N. caninum needs to invade the host cell in an active manner to survive. The increase in parasite cytosolic Ca2+ upon contact with the host cell mediates critical events, including the exocytosis of phylum-specific secretory organelles and the activation of the parasite invasion motor. Because invasion is considered a requirement for pathogen survival and replication within the host, the identification of secreted proteins (secretome) involved in invasion may be useful to reveal interesting targets for therapeutic intervention.\nMETHODS: To chart the currently missing N. caninum secretome, we employed mass spectrometry-based proteomics to identify proteins present in the N. caninum tachyzoite using two different approaches. The first approach was identifying the proteins present in the tachyzoite-secreted fraction (ESA). The second approach was determining the relative quantification through peptide stable isotope labelling of the tachyzoites submitted to an ethanol secretion stimulus (discharged tachyzoite), expecting to identify the secreted proteins among the down-regulated group.\nRESULTS: As a result, 615 proteins were identified at ESA and 2,011 proteins quantified at the discharged tachyzoite. We have analysed the connection between the secreted and the down-regulated proteins and searched for putative regulators of the secretion process among the up-regulated proteins. An interaction network was built by computational prediction involving the up- and down-regulated proteins. The mass spectrometry proteomics data have been deposited to the ProteomeXchange with identifier PXD000424.\nCONCLUSIONS: The comparison between the protein abundances in ESA and their measure in the discharged tachyzoite allowed for a more precise identification of the most likely secreted proteins. Information from the network interaction and up-regulated proteins was important to recognise key proteins potentially involved in the metabolic regulation of secretion. Our results may be helpful to guide the selection of targets to be investigated against Neospora caninum and other Apicomplexan organisms.", "claims": [{"id": "claim-data-access", "statement": "The mass spectrometry proteomics data have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository [83] with the dataset identifier PXD000424.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:24267406", "locator": "sections[28] \"Associated content\" paragraph 2", "quote": "The mass spectrometry proteomics data have been deposited in the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository [83] with the dataset identifier PXD000424."}], "state": "complete"}, {"id": "claim-target-result", "statement": "Using ESA proteomics and stable-isotope-labeling relative quantification after ethanol-triggered secretion, the study identified 615 ESA proteins and quantified 2,011 discharged-tachyzoite proteins; the source data were deposited in ProteomeXchange under PXD000424.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:24267406", "locator": "sections[0] \"Abstract\" paragraph 2", "quote": "The first approach was identifying the proteins present in the tachyzoite-secreted fraction (ESA). The second approach was determining the relative quantification through peptide stable isotope labelling of the tachyzoites submitted to an ethanol secretion stimulus (discharged tachyzoite), expecting to identify the secreted proteins among the down-regulated group."}, {"asset": "source:pmid:24267406", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "As a result, 615 proteins were identified at ESA and 2,011 proteins quantified at the discharged tachyzoite."}, {"asset": "source:pmid:24267406", "locator": "sections[24] \"Comparison of both approaches\" paragraph 2", "quote": "This could indicate that the fold change threshold of 2 might be too strict, since the mechanism of protein release from the three organelles is still not completely understood."}, {"asset": "source:pmid:24267406", "locator": "sections[0] \"Abstract\" paragraph 3", "quote": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange with identifier PXD000424."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "Using ESA proteomics and stable-isotope-labeling relative quantification after ethanol-triggered secretion, the study identified 615 ESA proteins and quantified 2,011 discharged-tachyzoite proteins; the source data were deposited in ProteomeXchange under PXD000424.", "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:24267406"], "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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