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Assessment of the role of heparan sulfate in high molecular weight kininogen binding to human umbilical vein endothelial cells
Author(s) -
Fernando L.P.,
Fernando A.N.,
Joseph K.,
Kaplan A.P.
Publication year - 2003
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1046/j.1538-7836.2003.00438.x
Subject(s) - heparan sulfate , high molecular weight kininogen , chemistry , umbilical vein , perlecan , kininogen , microbiology and biotechnology , endothelial stem cell , biochemistry , human umbilical vein endothelial cell , heparin , kallikrein , biology , in vitro , enzyme
Summary. The assembly and activation of the kinin forming system components on human umbilical vein endothelial cells (HUVEC) have been studied in great detail. Proteins such as gC1qR, cytokeratin‐1 and u‐PAR have been identified to be responsible for Zn 2+ ‐dependent binding of high molecular weight kininogen (HK) to HUVEC. Heparan sulfate has also been shown to have a major role in Zn 2+ ‐dependent binding of HK to the endothelial cell line, Ea.hy 926. In this study, we have analyzed the possible contribution of heparan sulfate to high molecular weight kininogen binding to HUVEC using multiple approaches. The presence of heparan sulfate on HUVEC was analyzed by staining with an antibody specific for heparan sulfate. Incubation of the cells with bacterial heparinases removed the heparan sulfate from the cell surface to the level seen with a control antibody, however, the Zn 2+ ‐dependent binding of HK was not affected. Further, blocking of heparan sulfate with a specific antibody to heparan sulfate even after digestion with heparinases did not reduce HK binding whereas antibodies to the proteins gC1qR and cytokeratin‐1 consistently reduced the binding of HK to the endothelial cells. The binding intensities of FITC‐labeled HK were similar in heparinase‐treated and ‐untreated HUVEC. The rate of kallikrein formation by the assembly of factor XII, HK and PK were similar in both heparinase‐treated and non‐treated HUVEC. All of these data indicate that heparan sulfate does not contribute significantly to HK binding to HUVEC.