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

Detalhes bibliográficos
Autor(a) principal: Pollo-Oliveira, Leticia
Data de Publicação: 2013
Outros Autores: Post, Harm, Acencio, Marcio Luis [UNESP], Lemke, Ney [UNESP], van den Toorn, Henk, Tragante, Vinicius, Heck, Albert J. R., Altelaar, A. F. Maarten, Yatsuda, Ana Patricia
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1186/1756-3305-6-335
http://hdl.handle.net/11449/111251
Resumo: 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.Methods: 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.Results: 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.Conclusions: 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.
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spelling Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomicsMass spectrometryNeospora caninumSecretomeShotgunRelative quantificationBackground: 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.Methods: 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.Results: 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.Conclusions: 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.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Netherlands Proteomics CentreCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Utrecht UniversityUniv Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Ribeirao Preto, SP, BrazilUniv Sao Paulo, Nucleo Apoio Pesquisa Prod Nat Sintet NPPNS, BR-14040903 Ribeirao Preto, SP, BrazilUniv Utrecht, Utrecht Inst Pharmaceut Sci, NL-3884 CH Utrecht, NetherlandsUniv Utrecht, Bijvoet Ctr Biomol Res, NL-3884 CH Utrecht, NetherlandsNetherlands Prote Ctr, NL-3884 CH Utrecht, NetherlandsUniv Estadual Paulista, UNESP, Botucatu Inst Biosci, BR-18918970 Botucatu, SP, BrazilUniv Med Ctr Utrecht, Dept Cardiol, Div Heart & Lungs, Utrecht, NetherlandsUniv Med Ctr Utrecht, Dept Med Genet, Div Biomed Genet, Utrecht, NetherlandsUniv Estadual Paulista, UNESP, Botucatu Inst Biosci, BR-18918970 Botucatu, SP, BrazilFAPESP: 05/53785-9FAPESP: 10/20684-3FAPESP: 13/02018-4CNPq: 480039/2009-7Biomed Central Ltd.Universidade de São Paulo (USP)Univ UtrechtNetherlands Prote CtrUniversidade Estadual Paulista (Unesp)Univ Med Ctr UtrechtPollo-Oliveira, LeticiaPost, HarmAcencio, Marcio Luis [UNESP]Lemke, Ney [UNESP]van den Toorn, HenkTragante, ViniciusHeck, Albert J. R.Altelaar, A. F. MaartenYatsuda, Ana Patricia2014-12-03T13:07:06Z2014-12-03T13:07:06Z2013-11-23info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article14application/pdfhttp://dx.doi.org/10.1186/1756-3305-6-335Parasites & Vectors. London: Biomed Central Ltd, v. 6, 14 p., 2013.1756-3305http://hdl.handle.net/11449/11125110.1186/1756-3305-6-335WOS:000328835300001WOS000328835300001.pdf7977035910952141Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengParasites & Vectors3.1631,702info:eu-repo/semantics/openAccess2023-12-30T06:18:19Zoai:repositorio.unesp.br:11449/111251Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T21:41:32.852981Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
title Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
spellingShingle Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
Pollo-Oliveira, Leticia
Mass spectrometry
Neospora caninum
Secretome
Shotgun
Relative quantification
title_short Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
title_full Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
title_fullStr Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
title_full_unstemmed Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
title_sort Unravelling the Neospora caninum secretome through the secreted fraction (ESA) and quantification of the discharged tachyzoite using high-resolution mass spectrometry-based proteomics
author Pollo-Oliveira, Leticia
author_facet Pollo-Oliveira, Leticia
Post, Harm
Acencio, Marcio Luis [UNESP]
Lemke, Ney [UNESP]
van den Toorn, Henk
Tragante, Vinicius
Heck, Albert J. R.
Altelaar, A. F. Maarten
Yatsuda, Ana Patricia
author_role author
author2 Post, Harm
Acencio, Marcio Luis [UNESP]
Lemke, Ney [UNESP]
van den Toorn, Henk
Tragante, Vinicius
Heck, Albert J. R.
Altelaar, A. F. Maarten
Yatsuda, Ana Patricia
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade de São Paulo (USP)
Univ Utrecht
Netherlands Prote Ctr
Universidade Estadual Paulista (Unesp)
Univ Med Ctr Utrecht
dc.contributor.author.fl_str_mv Pollo-Oliveira, Leticia
Post, Harm
Acencio, Marcio Luis [UNESP]
Lemke, Ney [UNESP]
van den Toorn, Henk
Tragante, Vinicius
Heck, Albert J. R.
Altelaar, A. F. Maarten
Yatsuda, Ana Patricia
dc.subject.por.fl_str_mv Mass spectrometry
Neospora caninum
Secretome
Shotgun
Relative quantification
topic Mass spectrometry
Neospora caninum
Secretome
Shotgun
Relative quantification
description 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.Methods: 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.Results: 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.Conclusions: 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.
publishDate 2013
dc.date.none.fl_str_mv 2013-11-23
2014-12-03T13:07:06Z
2014-12-03T13:07:06Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1186/1756-3305-6-335
Parasites & Vectors. London: Biomed Central Ltd, v. 6, 14 p., 2013.
1756-3305
http://hdl.handle.net/11449/111251
10.1186/1756-3305-6-335
WOS:000328835300001
WOS000328835300001.pdf
7977035910952141
url http://dx.doi.org/10.1186/1756-3305-6-335
http://hdl.handle.net/11449/111251
identifier_str_mv Parasites & Vectors. London: Biomed Central Ltd, v. 6, 14 p., 2013.
1756-3305
10.1186/1756-3305-6-335
WOS:000328835300001
WOS000328835300001.pdf
7977035910952141
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Parasites & Vectors
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1,702
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eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 14
application/pdf
dc.publisher.none.fl_str_mv Biomed Central Ltd.
publisher.none.fl_str_mv Biomed Central Ltd.
dc.source.none.fl_str_mv Web of Science
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
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