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A possible approach for gel-based proteomic studies in recalcitrant woody plants

2013-05-08Journal article Open access
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dc.contributor.authorSebastiana, Mónica
dc.contributor.authorFigueiredo, Andreia
dc.contributor.authorMonteiro, Filipa
dc.contributor.authorMartins, Joana
dc.contributor.authorFranco, Catarina
dc.contributor.authorCoelho, Ana Varela
dc.contributor.authorVaz, Fátima
dc.contributor.authorSimões, Tânia
dc.contributor.authorPenque, Deborah
dc.contributor.authorPais, Maria Salomé
dc.contributor.authorFerreira, Silvia
dc.date.accessioned2013-09-25T13:16:12Z
dc.date.available2013-09-25T13:16:12Z
dc.date.issued2013-05-08
dc.description.abstractWoody plants are particularly difficult to investigate due to high phenolic, resin, and tannin contents and laborious sample preparation. In particular, protein isolation from woody plants for two-dimensional gel electrophoresis (2-DE) is challenging as secondary metabolites negatively interfere with protein extraction and separation. In this study, three protein extraction protocols, using TCA, phenol and ethanol as precipitation or extraction agents, were tested in order to select the more efficient for woody recalcitrant plant gel-based proteomics. Grapevine leaves, pine needles and cork oak ectomycorrhizal roots were used to represent woody plant species and tissues. The phenol protocol produced higher quality 2-DE gels, with increased number of resolved spots, better spot focusing and representation of all molecular mass and isoelectric point ranges tested. In order to test the compatibility of the phenol extracted proteomes with protein identification several spots were excised from the phenol gels and analyzed by mass spectrometry (MALDI-TOF/TOF). Regardless the incomplete genome/protein databases for the plant species under analysis, 49 proteins were identified by Peptide Mass Fingerprint (PMF). Proteomic data have been deposited to the ProteomeXchange with identifier PXD000224. Our results demonstrate the complexity of protein extraction from woody plant tissues and the suitability of the phenol protocol for obtaining high quality protein extracts for efficient 2-DE separation and downstream applications such as protein identification by mass spectrometry.por
dc.identifier.citationSpringerplus. 2013 May 8;2(1):210. Print 2013 Dec.por
dc.identifier.issn2193-1801
dc.identifier.otherdoi:10.1186/2193-1801-2-210
dc.identifier.urihttp://hdl.handle.net/10400.18/1730
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherSpringerOpenpor
dc.relation.publisherversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3663981/por
dc.subjectGenómica Funcional e Estruturalpor
dc.subjectGrapevinepor
dc.subjectPinepor
dc.subjectOakpor
dc.subjectEctomycorrhizal Rootspor
dc.subjectProtein Extractionpor
dc.subject2-DEpor
dc.subjectMass Spectrometrypor
dc.titleA possible approach for gel-based proteomic studies in recalcitrant woody plantspor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage210por
oaire.citation.startPage210por
oaire.citation.volume2(1)por
rcaap.rightsopenAccesspor
rcaap.typearticlepor

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