PXD000244

PXD000244 is an original dataset announced via ProteomeXchange.

Title	14-3-3 membrane protein interactome
Description	To elucidate more details of the interaction between the PM (plasma membrane) H+ ATPase and 14-3-3 and to identify novel interaction partners of this multi-protein complex we developed a strategy for identification of protein-protein interactions using /in vivo/ formaldehyde cross-linking in combination with immunoprecipitation and MS-based protein identification. This method identified numerous proteins which may interact with the PM H+ ATPase to regulate its activity or to transmit environmental signals across the plasma membrane. All tryptic peptides mixtures were analyzed by LC-MS/MS using a nanoflow Easy-nLC System (Thermo Scientific, http://www.thermo.com) and an LTQ-Orbitrap hybrid mass spectrometer(Thermo Scientific) as a mass analyser. Peptides were separated on a 75 µm C18 analytical column (Integrafrit Biobasic, New Objectives, USA) with a linear gradient (10 to 64% acetonitrile) and sprayed directly into the LTQ-Orbitrap mass spectrometer. Data processing: Fragmentation spectra were searched against the NCBI non-redundant protein database (www.ncbi.nlm.nih.gov) with taxonomic restriction to all plant species using the Mascot software (version 2.2, Matrix Science, UK; www.matrixscience.com ) with the following search parameters: trypsin as cleavage enzyme, peptide mass tolerance 300 ppm, MS/MS tolerance 0.8 Da, cysteine carbamidomethylation as fixed modification, methionine oxidation as variable modification. Only peptides longer than 5 amino acids were considered. Peptides were accepted automatically if they displayed a Mascot score higher than 47 which was the Mascot significance score threshold (p < 0.01). False positive rate as estimated from a search against the decoy NCBI database was 1.34%.
HostingRepository	PRIDE
AnnounceDate	2016-05-10
AnnouncementXML	Submission_2016-05-10_00:16:49.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Waltraud Schulze
SpeciesList	scientific name: Lilium; NCBI TaxID: 4688;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2013-05-07 13:35:13	ID requested
⏵ 1	2016-05-10 00:16:51	announced

Pertl-Obermeyer H, Schulze WX, Obermeyer G, In vivo cross-linking combined with mass spectrometry analysis reveals receptor-like kinases and Ca(2+) signalling proteins as putative interaction partners of pollen plasma membrane H(+) ATPases. J Proteomics, 108():17-29(2014) [pubmed]

submitter keyword: plasma membrane proton ATPase, interaction partners, cross linking, 14-3-3 protein, LC-MS/MS

Waltraud Schulze
contact affiliation	Universität Hohenheim, Department of Plant Systems Biology
contact email	wschulze@uni-hohenheim.de
lab head
Waltraud Schulze
contact affiliation	Department of Plant Systems Biology
contact email	wschulze@uni-hohenheim.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD000244
     1. Label: PRIDE project
     2. Name: 14-3-3 membrane protein interactome