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PXD009965

PXD009965 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleTitle: Identification of LEFKOTHEA/PORR protein partners
DescriptionLEFKOTHEA (At5g62990) is a nuclear gene encoding an RNA-binding protein that carries a transit peptide (TP) and monopartite or bipartite nuclear localization signals (NLS). These motifs result to dual-targeting of LEFKOTHEA to the nucleus and chloroplasts. To identify the chloroplast and nuclear protein interactors of LEFKOTHEA, co-immunoprecipitation coupled to mass spectrometry (co-IP/MS) analysis was performed. The construct of LEFKOTHEA gene, expressed under the control of the endogenous promoter, with a FLAG tag at the C-terminus was introduced in Arabidopsis lefko2 mutant plants using the Agrobacterium system. The proteins were extracted under native conditions and incubated with anti-FLAG M2-agarose affinity gel. High salt concentrations and 3X FLAG peptide were applied to the beads for protein elution. The eluted protein fractions were subjected to Ultra High Pressure nanoLC-MS/MS. The LEFKOTHEA protein partners were identified using the MaxQuant software (version 1.5.3.30) against the complete Uniprot proteome of Arabidopsis thaliana (version of 19/1/2016) and a common contaminants database by the Andromeda search engine. Protein abundance was calculated on the basis of the normalized spectral protein intensity as label free quantitation (LFQ intensity). The statistical analysis was performed using Perseus (version 1.5.3.2).
HostingRepositoryPRIDE
AnnounceDate2019-08-08
AnnouncementXMLSubmission_2019-08-08_06:17:17.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterMartina Samiotaki
SpeciesList scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationListdeaminated residue; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
InstrumentLTQ Orbitrap
Dataset History
RevisionDatetimeStatusChangeLog Entry
02018-05-30 05:52:28ID requested
12019-08-08 06:17:18announced
Publication List
Dataset with its publication pending
Keyword List
curator keyword: Biological
submitter keyword: Lefkothea, PORR, Arabidopsis Thaliana. Affinity purification, interactors, endogeneous
Contact List
Polydefkis Hatzopoulos
contact affiliationDepartment of Biotechnology, Agricultural University of Athens, Iera Odos 75, Athens 118 55, Greece
contact emailphat@aua.gr
lab head
Martina Samiotaki
contact affiliationProtein Analysis Laboratory B.S.R.C. "Alexander Fleming", Alexander Fleming Street 34 16672, Vari, Greece
contact emailsamiotaki@fleming.gr
dataset submitter
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Dataset FTP location
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