Please use this identifier to cite or link to this item: https://hdl.handle.net/10316/107322
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dc.contributor.authorAnjo, Sandra I.-
dc.contributor.authorMelo, Matilde N.-
dc.contributor.authorLoureiro, Liliana R.-
dc.contributor.authorSabala, Lúcia-
dc.contributor.authorCastanheira, Pedro-
dc.contributor.authorGrãos, Mário-
dc.contributor.authorManadas, Bruno-
dc.date.accessioned2023-07-03T11:09:16Z-
dc.date.available2023-07-03T11:09:16Z-
dc.date.issued2019-04-
dc.identifier.issn22132317pt
dc.identifier.urihttps://hdl.handle.net/10316/107322-
dc.description.abstractMost of the redox proteomics strategies are focused on the identification and relative quantification of cysteine oxidation without considering the variation in the total levels of the proteins. However, protein synthesis and protein degradation also belong to the regulatory mechanisms of the cells, being therefore important to consider the changes in total protein levels in PTMs-focused analyses, such as cysteine redox characterization. Therefore, a novel integrative approach combining the SWATH-MS method with differential alkylation using a combination of commonly available alkylating reagents (oxSWATH) is presented, by which it is possible to integrate the information regarding relative cysteine oxidation with the analysis of the total protein levels in a cost-effective high-throughput approach. The proposed method was tested using a redox-regulated protein and further applied to a comparative analysis of secretomes obtained from cells cultured under control or oxidative stress conditions to strengthen the importance of considering the overall proteome changes. Using the OxSWATH method it was possible to determine both the relative proportion of reduced and reversible oxidized oxoforms, as well as the total levels of each oxoform by taking into consideration the total levels of the protein. Therefore, using OxSWATH the comparative analyses can be performed at two different levels by considering the relative proportion or the total levels at both peptide and protein level. Moreover, since samples are acquired in SWATH-MS mode, besides the redox centered analysis, a generic differential protein expression analysis can also be performed, allowing a truly comprehensive evaluation of proteomics changes upon the oxidative stimulus. Data are available via ProteomeXchange and SWATHAtlas with the identifiers PXD006802, PXD006802, and PASS01210.pt
dc.description.sponsorshipThis work was financed by the European Regional Development Fund (ERDF) through the COMPETE 2020 – Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT – Fundação para a Ciência e a Tecnologia, I.P., under projects: PTDC/NEU-NMC/0205/2012, POCI-01-0145-FEDER-007440 (ref.; UID/NEU/04539/2013), POCI-01-0145-FEDER-016428 (ref.: SAICTPAC/0010/2015), POCI-01-0145-FEDER-016795 (ref.: PTDC/ NEU-SCC/7051/2014), POCI-01-0145-FEDER-029311 (ref.: PTDC/ BTM-TEC/29311/2017), POCI-01-0145-FEDER-30943 (ref.: PTDC/ MEC-PSQ/30943/2017) and PTDC/MED-NEU/27946/2017; and by The National Mass Spectrometry Network (RNEM) under the contract POCI-01-0145-FEDER-402-022125 (ref.: ROTEIRO/0028/2013). S.I.A, was supported by Ph.D. fellowship SFRH/BD/81495/2011, co-financed by the European Social Fund (ESF) through the POCH – Programa Operacional do Capital Humano and national funds via FCT.pt
dc.language.isoengpt
dc.publisherElsevierpt
dc.relationPTDC/NEU-NMC/0205/2012pt
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID/NEU/04539/2013pt
dc.relationSAICTPAC/0010/2015pt
dc.relationPTDC/NEU-SCC/7051/2014pt
dc.relationPTDC/MED-NEU/27946/2017pt
dc.rightsopenAccesspt
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/pt
dc.titleoxSWATH: An integrative method for a comprehensive redox-centered analysis combined with a generic differential proteomics screeningpt
dc.typearticle-
degois.publication.firstPage101130pt
degois.publication.titleRedox Biologypt
dc.peerreviewedyespt
dc.identifier.doi10.1016/j.redox.2019.101130pt
degois.publication.volume22pt
dc.date.embargo2019-04-01*
uc.date.periodoEmbargo0pt
item.grantfulltextopen-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.fulltextCom Texto completo-
item.openairetypearticle-
item.cerifentitytypePublications-
item.languageiso639-1en-
crisitem.author.researchunitCNC - Center for Neuroscience and Cell Biology-
crisitem.author.researchunitCNC - Center for Neuroscience and Cell Biology-
crisitem.author.researchunitCNC - Center for Neuroscience and Cell Biology-
crisitem.author.orcid0000-0002-2707-1488-
crisitem.author.orcid0000-0002-2087-4042-
crisitem.project.grantnoCNC. IBILI-
crisitem.project.grantnoMEDPERSYS - Synaptic networks and Personalized Medicine Approaches to Understand Neurobehavioural Diseases Across the Lifespan-
crisitem.project.grantnoSCZ_bioMark Schizophrenia diagnosis and prognosis: finding the way to a personalized medicine-
Appears in Collections:FMUC Medicina - Artigos em Revistas Internacionais
I&D CNC - Artigos em Revistas Internacionais
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