PXD047399

PXD047399 is an original dataset announced via ProteomeXchange.

Title	Endothelial regulation of platelet activation and signaling: endothelial effects on the platelet phosphoproteome
Description	The endothelial regulation of platelet activity is incompletely understood. We investigated how the presence of human umbilical vein endothelial cells (HUVEC) in whole-blood microfluidics affected platelet activation induced by collagen receptor glycoprotein VI (GPVI) and the PAR receptors for thrombin. On collagen/tissue factor surface, HUVEC potently suppressed platelet adhesion and Ca2+ rises upon high-shear blood flow at physiological temperature. HUVEC accordingly suppressed thrombus and fibrin formation. Similarly under stasis, platelet exposure to HUVEC (1-30 min) reduced Ca2+ responses to collagen-related peptide (CRP-XL, GPVI agonist) and thrombin (PAR agonist). Parallel samples of platelets from 3 donors, exposed to HUVEC, CRP-XL and/or thrombin, were analyzed by label-free phosphoproteome analysis. High-resolution mass spectrometry gave 5,463 platelet phosphopeptides, corresponding to 1,472 proteins, with good correlation between biological and technical replicates (R>0.86). Stringent filtering steps revealed 26 principal pathways (Reactome) and 143 kinase substrates (KEGG, PhosphoSitePlus). Repeated phosphoproteome analysis indicated a set of protein phosphorylation sites that was differentially (44) or similarly (110) regulated by HUVEC exposure or by agonist stimulation. The differential regulation was confirmed by stable-isotope analysis of platelets from 2 additional donors. Substrate analysis indicated preferential involvement of MAPK, CDK, DYRK, STK and PKC protein-kinase classes. Collectively, our results reveal a resetting of the protein phosphorylation profile in platelets exposed to endothelium or to conventional agonists, and to the endothelium-promoted activation of a multi-kinase network, beyond classical prostacyclin and nitric oxide actors, that may contribute to platelet inhibition.
HostingRepository	PRIDE
AnnounceDate	2026-09-08
AnnouncementXML	Submission_2026-09-08_06:30:05.816.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Fiorella Andrea Solari
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	phosphorylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF; Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-11-30 05:07:41	ID requested
⏵ 1	2026-09-08 06:30:06	announced

10.1096/fj.202300360rr;

Provenzale I, Solari FA, Sch, ö, nichen C, Brouns SLN, Fern, á, ndez DI, Kuijpers MJE, van der Meijden PEJ, Gibbins JM, Sickmann A, Jones C, Heemskerk JWM, Endothelium-mediated regulation of platelet activation: Involvement of multiple protein kinases. FASEB J, 38(4):e23468(2024) [pubmed]

submitter keyword: phosphoproteomcis,platelets

Albert Sickmann
contact affiliation	Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V.
contact email	albert.sickmann@isas.de
lab head
Fiorella Andrea Solari
contact affiliation	Leibniz-Institut für Analytische Wissenschaften – ISAS – e.V.
contact email	fiorella.solari@isas.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/2026/09/PXD047399

PRIDE project URI

• PRIDE
  1. PXD047399
     1. Label: PRIDE project
     2. Name: Endothelial regulation of platelet activation and signaling: endothelial effects on the platelet phosphoproteome