PXD001887

PXD001887 is an original dataset announced via ProteomeXchange.

Title	Deep, quantitative coverage of the lysine acetylome using novel anti-acetyl-lysine antibodies
Description	Svinkina T, Gu H, Silva JC, Mertins P, Qiao J, Fereshetian S, Jaffe JD, Kuhn E, Udeshi ND,Carr SA. Mol Cell Proteomics, 2015. Introduction of antibodies specific for acetylated lysine has significantly improved the detection of endogenous acetylation sites by mass spectrometry. Here, we describe a new, commercially available mixture of anti-Kac antibodies and show its utility for in-depth profiling of the acetylome. Specifically, seven complementary monoclones with high specificity for Kac were combined into a final anti-Kac reagent which results in at least a 2-fold increase in identification of Kac peptides over a commonly used Kac antibody. We outline optimal antibody usage conditions, effective offline bRP separation, and use of state-of-the art LC-MS technology for achieving unprecedented coverage of the acetylome. The methods presented in this study enabled the quantification of over 10,000 Kac peptides from over 3000 Kac proteins from a single SILAC labeled sample using 7.5 mg of peptide input per state. These methods result in the deepest coverage of acetylation sites from SAHA treated Jurkat cells. The approach was also applied to breast tumor xenograft samples using isobaric mass tag labeling of peptides (iTRAQ4-plex, TMT6 and TMT10-plex reagents) for quantification. Greater than 6,700 Kac peptides from over 2,300 Kac proteins were quantified using 1 mg of tumor per iTRAQ 4-plex channel.
HostingRepository	MassIVE
AnnounceDate	2015-10-02
AnnouncementXML	Submission_2015-10-02_20:23:37.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Karl Clauser
SpeciesList	scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606; scientific name: Mus musculus; common name: house mouse; NCBI TaxID: 10090;
ModificationList	Acetyl; TMT6plex; unknown modification: TMT10plex; iTRAQ4plex; Carbamidomethyl
Instrument	Q Exactive; LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2015-03-08 20:06:10	ID requested
⏵ 1	2015-10-02 20:23:38	announced

Dataset with its publication pending

submitter keyword: lysine acetylome

Steven A. Carr
lab head
Karl Clauser
contact affiliation	Broad Institute of MIT and Harvard
contact email	clauser@broad.mit.edu
dataset submitter

MassIVE dataset URI

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