PXD000597

PXD000597 is an original dataset announced via ProteomeXchange.

Title	Quantitative phosphoproteomics unveils temporal dynamics of thrombin signaling in human endothelial cells
Description	Thrombin is the key serine protease of the coagulation cascade and a potent trigger of protease-activated receptor (PAR)1-mediated platelet aggregation. In recent years, PAR1 has become an appealing target for anticoagulant therapies. However, the inhibitors that have been developed so far increase bleeding risk in patients, likely because they interfere with endogenous PAR1 signaling in the endothelium. Due to its complexity, thrombin-induced signaling in endothelial cells has remained incompletely understood. Here, we have combined stable isotope amino acids in cell culture, affinity-based phospho-peptide enrichment and high resolution mass spectrometry and performed a time-resolved analysis of the thrombin-induced signaling in human primary endothelial cells. We identified 2224 thrombin-regulated phosphorylation sites, the majority of which has not been previously related to thrombin. Those sites were localized on proteins which are novel to thrombin signaling, but also on well-known players such as PAR1, Rho-associated kinase 2, phospholipase C, and proteins related to actin cytoskeleton, cell-cell junctions and Weibel-Palade body release. Our study provides a unique resource of phosphoproteins and phosphorylation sites which may generate novel insights into an intimate understanding of thrombin-mediated PAR signaling and the development of improved PAR1 antagonists that affect platelet but not endothelial cell function.
HostingRepository	PRIDE
AnnounceDate	2014-07-24
AnnouncementXML	Submission_2014-07-24_03:55:55.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sara Zanivan
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; acetylated residue; phosphorylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2013-11-22 05:27:17	ID requested
1	2014-02-07 05:21:26	announced
⏵ 2	2014-07-24 03:55:56	announced	Updated project metadata.

van den Biggelaar M, Hern, á, ndez-Fernaud JR, van den Eshof BL, Neilson LJ, Meijer AB, Mertens K, Zanivan S, Quantitative phosphoproteomics unveils temporal dynamics of thrombin signaling in human endothelial cells. Blood, 123(12):e22-36(2014) [pubmed]

curator keyword: Biomedical

submitter keyword: Thrombin, PAR, endothelial cells, phosphoproteomics, secretome, SILAC

Sara Zanivan
contact affiliation	CRUK Beatson Institute
contact email	s.zanivan@beatson.gla.ac.uk
lab head
Sara Zanivan
contact affiliation	Vascular Proteomics
contact email	s.zanivan@beatson.gla.ac.uk
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/2014/02/PXD000597

PRIDE project URI

• PRIDE
  1. PXD000597
     1. Label: PRIDE project
     2. Name: Quantitative phosphoproteomics unveils temporal dynamics of thrombin signaling in human endothelial cells