Binding of factors IX and IXa to cultured vascular endothelial cells.
Factor IX and its activated form IXa have been found to bind to confluent cultured bovine aortic and human umbilical vein endothelial cells. Binding of bovine factors IX and IXa to the bovine endothelial cells was saturable and specific and reached a plateau in 75 min at 4 degrees C and 30 min at 37 degrees C. Binding was half-maximal at a total factor IX or IXa concentration of 2.3 +/- 0.2 nM. At 4 degrees C, a maximum of 42 fmol of tritiated factor IX or IXa bound to 10(6) cells (an average of 20,000 molecules per cell). The binding of tritiated factor IX or IXa was inhibited by excess unlabeled factor IX or IXa but not by factor X, prothrombin, or thrombin. Competition studies indicated that factors IX and IXa interacted with the same site. Binding was reversible, with 50% of the specifically bound factor IX or IXa eluted in 40 min by a 400-fold excess of unlabeled protein. Specific binding required Ca2+ with half-maximal binding at 1.2 mM CaCl2. Factor IXa bound to the cells was tested for procoagulant activity in a clotting assay with factor IX-deficient plasma, cephalin, and CaCl2. Cell-bound factor IXa was at least 3-fold more active than was factor IXa in solution. The retention of procoagulant activity by cell surface-bound factor IXa provides a mechanism for the localization of clot-promoting activity.
D000818:Animals; D001011:Aorta / Q000502:physiology*; D001667:Binding, Competitive; D001779:Blood Coagulation Factors / Q000378:metabolism*; D002417:Cattle; D004727:Endothelium / Q000502:physiology; D005164:Factor IX / Q000378:metabolism*; D015949:Factor IXa; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D013045:Species Specificity
D001779:Blood Coagulation Factors; D005164:Factor IX; D015949:Factor IXa