RESEARCH ARTICLE
Analysis of Endothelial Protein C Receptor Functionality on Living Cells’
E Ducros1, S Mirshahi2, C Bermot1, M Mirshahi*, 1
Article Information
Identifiers and Pagination:
Year: 2009Volume: 3
First Page: 49
Last Page: 54
Publisher ID: TOBIOCJ-3-49
DOI: 10.2174/1874091X00903010049
Article History:
Received Date: 11/2/2009Revision Received Date: 11/3/2009
Acceptance Date: 20/3/2009
Electronic publication date: 13/5/2009
Collection year: 2009
open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.
Abstract
Activated protein C (APC) is a major control system of blood coagulation. APC prevents coagulation pathway by degrading Va and VIIIa plasma’s coagulation factors. Protein C activation requires its binding to specific endothelial cell receptor (EPCR). APC binding to EPCR also activates a wide range of defense mechanisms (anti-inflammatory, antiapoptosis…). EPCR expression by cells can be detected by various methods, including immunoanalysis and molecular biology. However, no assays evaluate its functionality. A method, inspired of a standard fibrinoformation time assay, was developed to estimate EPCR ability to bind APC on living cell surface in vitro. Endothelial cells were incubated with APC and fibrinoformation on cells was followed by spectrophotometry (plasma absorbance increases with fibrin polymerization). Membrane-bound EPCR retain APC, thus prolonging fibrinoformation time in a dose-dependent manner. Control was realized with EPCR-negative cells. This new method can be used on any cell type to study the expression of other coagulation receptors.
