PXD000543

PXD000543 is an original dataset announced via ProteomeXchange.

Title	Quantitative analysis of murine T-cells
Description	1D-LC-MS/MS analysis of OGE-prefractionated and TMT labeled mouse samples consiting of 2 unstimulated and 2 stimulated T-cell samples. The acquired raw-files were converted to the mascot generic file (mgf) format using the msconvert tool (part of ProteoWizard, version 3.0.4624 (2013-6-3)). Using the MASCOT algorithm (Matrix Science, Version 2.4.0), the mgf files were searched against a decoy database containing normal and reverse sequences of the predicted SwissProt entries of mus musculus (www.ebi.ac.uk, release date 16/05/2012) and commonly observed contaminants (in total 33,832 sequences) generated using the SequenceReverser tool from the MaxQuant software (Version 1.0.13.13). The precursor ion tolerance was set to 10 ppm and fragment ion tolerance was set to 0.01 Da. The search criteria were set as follows: full tryptic specificity was required (cleavage after lysine or arginine residues unless followed by proline), 2 missed cleavages were allowed, carbamidomethylation (C), TMT6plex (K and peptide n-terminus) were set as fixed modification and oxidation (M) as a variable modification. Next, the database search results were imported to the Scaffold Q+ software (version 4.1.1, Proteome Software Inc., Portland, OR) and the protein false identification rate was set to 1% based on the number of decoy hits. Specifically, peptide identifications were accepted if they could be established at greater than 94.0% probability to achieve an FDR less than 1.0% by the scaffold local FDR algorithm. Protein identifications were accepted if they could be established at greater than 6.0% probability to achieve an FDR less than 1.0% and contained at least 1 identified peptide. Protein probabilities were assigned by the Protein Prophet program (Nesvizhskii, et al, Anal. Chem. 2003; 75(17):4646-58). Proteins that contained similar peptides and could not be differentiated based on MS/MS analysis alone were grouped to satisfy the principles of parsimony. Proteins sharing significant peptide evidence were grouped into clusters. For quantification, acquired reporter ion intensities in the experiment were globally normalized across all acquisition runs. Individual quantitative samples were normalized within each acquisition run. Intensities for each peptide identification were normalized within the assigned protein. The reference channels were normalized to produce a 1:1 fold change. All normalization calculations were performed using medians to multiplicatively normalize data. A list of identified and quantified proteins is available in the xls file.
HostingRepository	PRIDE
AnnounceDate	2014-12-05
AnnouncementXML	Submission_2014-12-05_00:30:24.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD000543
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Alexander Schmidt
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue; TMT6plex-126 reporter+balance reagent acylated residue
Instrument	LTQ Orbitrap; instrument model: LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2013-10-28 03:07:00	ID requested
1	2014-12-02 09:44:16	announced
⏵ 2	2014-12-05 00:30:25	announced	Updated project metadata.

Gruber AR, Martin G, M, ü, ller P, Schmidt A, Gruber AJ, Gumienny R, Mittal N, Jayachandran R, Pieters J, Keller W, van Nimwegen E, Zavolan M, Global 3' UTR shortening has a limited effect on protein abundance in proliferating T cells. Nat Commun, 5():5465(2014) [pubmed]

curator keyword: Biomedical, Biological

submitter keyword: T-cells, quantitative analysis, mouse, TMT labeling

Alexander Schmidt
contact affiliation	Proteomics Core Facility
contact email	alex.schmidt@unibas.ch
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/2014/12/PXD000543

PRIDE project URI

• PRIDE
  1. PXD000543
     1. Label: PRIDE project
     2. Name: Quantitative analysis of murine T-cells