The endothelial protein C receptor supports tissue factor ternary coagulation initiation complex signaling through protease-activated receptors

J Biol Chem. 2011 Feb 18;286(7):5756-67. doi: 10.1074/jbc.M110.201228. Epub 2010 Dec 13.

Abstract

Protease-activated receptor (PAR) signaling is closely linked to the cellular activation of the pro- and anticoagulant pathways. The endothelial protein C receptor (EPCR) is crucial for signaling by activated protein C through PAR1, but EPCR may have additional roles by interacting with the 4-carboxyglutamic acid domains of procoagulant coagulation factors VII (FVII) and X (FX). Here we show that soluble EPCR regulates the interaction of FX with human or mouse tissue factor (TF)-FVIIa complexes. Mutagenesis of the FVIIa 4-carboxyglutamic acid domain and dose titrations with FX showed that EPCR interacted primarily with FX to attenuate FX activation in lipid-free assay systems. In human cell models of TF signaling, antibody inhibition of EPCR selectively blocked PAR activation by the ternary TF-FVIIa-FXa complex but not by the non-coagulant TF-FVIIa binary complex. Heterologous expression of EPCR promoted PAR1 and PAR2 cleavage by FXa in the ternary complex but did not alter PAR2 cleavage by TF-FVIIa. In murine smooth muscle cells that constitutively express EPCR and TF, thrombin and FVIIa/FX but not FVIIa alone induced PAR1-dependent signaling. Although thrombin signaling was unchanged, cells with genetically reduced levels of EPCR no longer showed a signaling response to the ternary complex. These results demonstrate that EPCR interacts with the ternary TF coagulation initiation complex to enable PAR signaling and suggest that EPCR may play a role in regulating the biology of TF-expressing extravascular and vessel wall cells that are exposed to limited concentrations of FVIIa and FX provided by ectopic synthesis or vascular leakage.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / genetics
  • Antigens, CD / metabolism*
  • Cells, Cultured
  • Endothelial Protein C Receptor
  • Factor VIIa / genetics
  • Factor VIIa / metabolism
  • Factor X / genetics
  • Factor X / metabolism
  • Glycoproteins / genetics
  • Glycoproteins / metabolism*
  • Humans
  • Mice
  • Mice, Knockout
  • Multienzyme Complexes / genetics
  • Multienzyme Complexes / metabolism
  • Myocytes, Smooth Muscle / cytology
  • Myocytes, Smooth Muscle / metabolism*
  • Protein C / genetics
  • Protein C / metabolism
  • Protein Structure, Tertiary
  • Receptor, PAR-1 / genetics
  • Receptor, PAR-1 / metabolism*
  • Receptor, PAR-2 / genetics
  • Receptor, PAR-2 / metabolism*
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Signal Transduction / physiology*
  • Thrombin / genetics
  • Thrombin / metabolism
  • Thromboplastin / genetics
  • Thromboplastin / metabolism*

Substances

  • Antigens, CD
  • Endothelial Protein C Receptor
  • Glycoproteins
  • Multienzyme Complexes
  • PROCR protein, human
  • Procr protein, mouse
  • Protein C
  • Receptor, PAR-1
  • Receptor, PAR-2
  • Receptors, Cell Surface
  • Factor X
  • Thromboplastin
  • Factor VIIa
  • Thrombin