PXD000269

PXD000269 is an original dataset announced via ProteomeXchange.

Title	Minimalistic sample processing
Description	Proteomic sample preparation typically involves multi-step workflows that can potentially perturb the biological system under analysis. These datasets were used to demonstrate that the entire processing pipeline, from cell lysis through elution of purified peptides, can be performed in a single, enclosed volume. This in-StageTip sample preparation method largely eliminates contamination or losses and is extremely robust and scalable. Peptides can be eluted in several fractions or in one step for single-run proteomics. In one day of gradient time we obtained the largest proteomes for budding and fission yeast to date. Applying the in-StageTip method to quadruplicate measurements of a human cell line yielded copy number estimates for more than 9000 human proteins. Data Analysis: MS raw files were analyzed by MaxQuant software (version 1.3.10.12) and peak lists were searched either against the human Uniprot FASTA database version 2/25/2012 (81213 entries), against the S. cerevisiae Uniprot FASTA database version 2/25/2012 (6649 entries) or the SGD based S. cerevisiae Fasta database orf_trans.20100105 (5904 entries), or against the S. pombe Uniprot FASTA database version 4/2/2013 (5096 entries) or pompep FASTA database version 4/2/2013 (5031 entries) and a common contaminants database (247 entries) by Andromeda search engine with cysteine carbamidomethylation as a fixed modification and N-terminal acetylation and methionine oxidation as variable modifications. False discovery rate (FDR) was set to 0.01 for proteins and peptides (minimum length of 7 amino acids) and was determined by searching a reverse database. Enzyme specificity was set as C-terminal to Arg and Lys, and a maximum of 2 miscleavages allowed. Peptide identification was performed with an allowed initial precursor mass deviation up to 7 ppm and an allowed fragment mass deviation 20 ppm. Quantification of SILAC pairs was carried out by MaxQuant with standard settings and without the re-quantification option.
HostingRepository	PRIDE
AnnounceDate	2014-07-25
AnnouncementXML	Submission_2014-07-25_01:12:54.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Mario Oroshi
SpeciesList	scientific name: Schizosaccharomyces pombe (Fission yeast); NCBI TaxID: 4896; scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2013-05-28 15:42:05	ID requested
1	2014-01-31 11:49:34	announced
⏵ 2	2014-07-25 01:12:55	announced	Updated project metadata.

Dataset with its publication pending

submitter keyword: Yeast, in-depth proteome, human, copy number

Mario Oroshi
contact affiliation	Proteomics
contact email	oroshi@biochem.mpg.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/2014/01/PXD000269

PRIDE project URI

• PRIDE
  1. PXD000269
     1. Label: PRIDE project
     2. Name: Minimalistic sample processing