PXD000268

PXD000268 is an original dataset announced via ProteomeXchange.

Title	AKT isoform specific signaling
Description	Phosphoproteome analysis of HL-1 cardiomyocytes carrying AKT1 or AKT2 isoform specific knock down, respectively. Six biological replicates were analysed. Cells were grown until reaching 90 % confluency, starved for 50 minutes followed by stimulation with 200 nM insulin for 10 minutes. In solution stable isotope dimethyl labeling was performed following the protocol of Boersema et al. (2009) Nat. Protoc. 4, 484-494 Fe-NTA Phosphopeptide Enrichment Kit (Thermo Scientific) was used according to the manufacturer’s instructions. Peptides were desalted with Graphite Spin Columns (Thermo Scientific) and fractionated by automated off-line 2-D LC. First dimension: 1 mm × 15 cm Polysulfoethyl-Aspartamide column (Dionex, Thermo Scientific). Two replicates were alternatively fractionated in-solution using the Agilent 3100 OFFGEL Fractionator (24 cm IPG strips pH 3-10 (GE Healthcare) and a 24 well frame set from Agilent Technologies). Peptides from the single SCX fractions were further separated and analyzed by reversed-phase nano-LC-MS/MS. Sequence database-search of the MS data was performed against the uniprot mouse database (downloaded 13.06.2012) using MaxQuant (Version 1.3.0.5).
HostingRepository	PRIDE
AnnounceDate	2014-10-23
AnnouncementXML	Submission_2014-10-23_02:38:53.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Michael Reinartz
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue; phosphorylated residue; dimethylated residue
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2013-05-27 15:09:17	ID requested
1	2014-08-28 10:37:18	announced
⏵ 2	2014-10-23 02:38:55	announced	Updated project metadata.

Reinartz M, Raupach A, Kaisers W, G, ö, decke A, AKT1 and AKT2 induce distinct phosphorylation patterns in HL-1 cardiac myocytes. J Proteome Res, 13(10):4232-45(2014) [pubmed]

curator keyword: Biomedical, Biological, Cardiovascular

submitter keyword: HL-1 cardiac myocytes, AKT, phosphoproteome, LTQ-Orbitrap

Michael Reinartz
contact affiliation	Institut für Herz- und Kreislaufphysiologie
contact email	Michael.Reinartz@uni-duesseldorf.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2014/08/PXD000268

PRIDE project URI

• PRIDE
  1. PXD000268
     1. Label: PRIDE project
     2. Name: AKT isoform specific signaling