RT info:eu-repo/semantics/article
T1 Quantitative phosphoproteomics unveils temporal dynamics of thrombin signaling in human endothelial cells.
A1 Hernández Fernaud, Juan Ramón
A1 Biggelaar, Maartje van den
A1 Eshof, Bart L. van den
A1 Neilson, Lisa J.
A1 Meijer, Alexander B.
A1 Mertens, Koen
A1 Zanivan, Sara
A2 BioquímicaMicrobiología, Biología Celular y Genética
A2 The Beatson Institute for Cancer ResearchGlasgow, UK
AB Thrombin is the key serine protease of the coagulation cascade and a potent trigger of protease-activated receptor 1 (PAR1)-mediated platelet aggregation. In recent years, PAR1hasbecomeanappealingtargetforanticoagulanttherapies. However,theinhibitors that have been developed so far increase bleeding risk in patients, likely because they interfere with endogenous PAR1 signaling in the endothelium. Because of 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 phosphopeptide enrichment, and high-resolution mass spectrometry and performed a timeresolvedanalysisofthethrombin-inducedsignalinginhumanprimaryendothelialcells.We identified 2224 thrombin-regulated phosphorylation sites, the majority of which have not beenpreviouslyrelatedtothrombin.Thosesiteswerelocalizedonproteinsthatarenovelto thrombinsignaling,butalsoonwell-knownplayerssuchasPAR1,Rho-associatedkinase2, phospholipaseC,andproteinsrelatedtoactincytoskeleton,cell-celljunctions,andWeibelPalade body release. Our study provides a unique resource of phosphoproteins and phosphorylation sites that 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.
YR 2013
FD 2013
LK http://riull.ull.es/xmlui/handle/915/38826
UL http://riull.ull.es/xmlui/handle/915/38826
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DS Repositorio institucional de la Universidad de La Laguna
RD 02-ene-2025