Time-resolved characterization of cAMP/PKA-dependent signaling reveals that platelet inhibition is a concerted process involving multiple signaling pathways

  • Florian Beck
  • Jörg Geiger
  • Stepan Gambaryan
  • Johannes Veit
  • Marc Vaudel
  • Peter Nollau
  • Oliver Kohlbacher
  • Lennart Martens
  • Ulrich Walter
  • Albert Sickmann
  • René P Zahedi

Abstract

One of the most important physiological platelet inhibitors is endothelium-derived prostacyclin which stimulates the platelet cyclic adenosine monophosphate/protein kinase A (cAMP/PKA)-signaling cascade and inhibits virtually all platelet-activating key mechanisms. Using quantitative mass spectrometry, we analyzed time-resolved phosphorylation patterns in human platelets after treatment with iloprost, a stable prostacyclin analog, for 0, 10, 30, and 60 seconds to characterize key mediators of platelet inhibition and activation in 3 independent biological replicates. We quantified over 2700 different phosphorylated peptides of which 360 were significantly regulated upon stimulation. This comprehensive and time-resolved analysis indicates that platelet inhibition is a multipronged process involving different kinases and phosphatases as well as many previously unanticipated proteins and pathways.

Bibliographical data

Original languageEnglish
ISSN0006-4971
DOIs
Publication statusPublished - 30.01.2014
PubMed 24324209