Proteolysis of the endothelial cell protein C receptor by neutrophil proteinase 3
Author(s)
Type
Journal Article
Abstract
Background: The endothelial cell protein C receptor (EPCR) presents protein C to the thrombin:thrombomodulin complex on the endothelium of large vessels, and enhances the generation of activated protein C (APC) and activation of protease-activated receptor-1. A previous report has demonstrated binding of soluble (s) EPCR to activated neutrophils via surface proteinase 3 (PR3). Methods: We now report further characterization of this interaction. Activated neutrophils and purified PR3 both decrease endothelial cell (EC) surface EPCR, suggestive of its proteolysis. Results: When added to purified recombinant sEPCR, PR3 produced multiple cleavages, with early products including 20 kDa N-terminal and C-terminal (after Lys176) fragments. The binding of active site blocked PR3 to sEPCR was studied by surface plasmon resonance. Estimates of the KD of 18.5–102 nm were obtained with heterogeneous binding, suggestive of more than a single interaction site. Conclusions: This work demonstrates PR3 binding to and proteolysis of EPCR and suggests a mechanism by which anticoagulant and cell protective pathways can be down-regulated during inflammation.
Date Issued
2007-04-16
Date Acceptance
2007-02-19
Citation
Journal of Thrombosis and Haemostasis, 2007, 5 (5), pp.980-988
ISSN
1538-7933
Publisher
Wiley
Start Page
980
End Page
988
Journal / Book Title
Journal of Thrombosis and Haemostasis
Volume
5
Issue
5
Copyright Statement
© 2007 The Author(s). Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.
License URL
Subjects
Science & Technology
Life Sciences & Biomedicine
Hematology
Peripheral Vascular Disease
Cardiovascular System & Cardiology
HEMATOLOGY
PERIPHERAL VASCULAR DISEASE
endothelial cells
EPCR
inflammation
neutrophils
PR3
LARGE BLOOD-VESSELS
WEGENERS-GRANULOMATOSIS
ACTIVATED NEUTROPHILS
VASCULAR INJURY
PLASMA-LEVELS
IDENTIFICATION
EXPRESSION
THROMBOMODULIN
APOPTOSIS
Blood Coagulation Factors
Chromatography, High Pressure Liquid
Coculture Techniques
Flow Cytometry
Humans
Hydrolysis
Myeloblastin
Neutrophil Activation
Proteomics
Receptors, Cell Surface
Cardiovascular System & Hematology
1102 Cardiovascular Medicine And Haematology
1103 Clinical Sciences
Publication Status
Published