PXD018952-1

PXD018952 is an original dataset announced via ProteomeXchange.

Title	Lowering Sample Requirements to Study Tyrosine Kinase Signaling using Phosphoproteomics with the TMT Calibrator Approach
Description	Analysis of tyrosine kinase signaling is critical for the development of targeted cancer therapy. Currently, immunoprecipitation (IP) of phosphotyrosine (pY) peptides prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS) is used to profile kinase substrates. A typical protocol requests 10 mg of total protein from ~108 cells or 50-100 mg of tissue. Large sample requirements can be cost prohibitive or not feasible for certain experiments. Sample multiplexing in chemical labeling reduces the protein amount required for each sample, and newer approaches use a material-rich reference channel as a calibrator to trigger detection and quantification for even smaller samples. Here, we demonstrate that the tandem mass tag (TMT) calibrator approach reduces the sample input for pY profiling 10-fold (to ~1 mg total protein per sample from 107 cells grown in 1 plate), while maintaining the depth of pY proteome sampling and the biological content of the experiment. This strategy opens more opportunities for pY profiling of large sample cohorts and samples with limited protein quantity such as immune cells, xenograft models, and human tumors.
HostingRepository	PRIDE
AnnounceDate	2020-07-29
AnnouncementXML	Submission_2020-07-28_22:55:46.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD018952
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	John Koomen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	Phospho; TMT6plex; phosphorylated residue; Oxidation; Carbamidomethyl
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2020-05-01 17:40:58	ID requested
⏵ 1	2020-07-28 22:55:46	announced
2	2020-09-08 23:17:19	announced	2020-09-09: Updated publication reference for PubMed record(s): 32865326.

Dataset with its publication pending

submitter keyword: Tyrosine Phosphorylation, Cancer, Signaling, TMT, Calibrator

John M. Koomen, PhD
contact affiliation	Molecular Oncology Moffitt Cancer Center Tampa, FL, USA
contact email	john.koomen@moffitt.org
lab head
John Koomen
contact affiliation	Moffitt Cancer Center
contact email	john.koomen@moffitt.org
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD018952
     1. Label: PRIDE project
     2. Name: Lowering Sample Requirements to Study Tyrosine Kinase Signaling using Phosphoproteomics with the TMT Calibrator Approach