title: Interaction of C1 inhibitor with thrombin on the endothelial surface.
abstract: Thrombin, the central bioregulatory enzyme of haemostasis, also has a potent vasopermeability effect on the surface of endothelial cells, and has therefore been considered a major link between the activation of the coagulation pathway and inflammation. C1 inhibitor inhibits thrombin with a low second-order rate constant that can be increased by heparin. The aim of this study was to investigate whether the C1 inhibitor-induced inhibition of thrombin is potentiated on the endothelial surface. The interaction of C1 inhibitor and thrombin was evaluated in an in-vitro system of human umbilical vein endothelial cells (HUVECs) to which purified C1 inhibitor and thrombin have been added. The role of heparins and selectins has been tested by adding heparinase and Mab to selectins. Kinetic analysis under pseudo-first-order conditions showed that the inhibitory effect of C1 inhibitor on thrombin is greater on the surface of endothelial cells. After incubating nanomolar concentrations of thrombin and micromolar concentrations of C1 inhibitor in a purified system, thrombin activity remained significant, but was almost totally suppressed in the presence of HUVECs. The abolition of such suppression by heparinase and Mab to selectins supports the involvement of heparin and selectins in C1 inhibitor-thrombin interaction. Furthermore, the second-order rate constant was 25 ± 3 /s per mol/l in our purified system, but increased to 100 ± 9 /s per mol/l in the presence of HUVECs. Our results indicate that C1 inhibitor can inhibit thrombin activity on vascular endothelium via binding to selectins and potentiation by heparins. This may contribute to the modulation of thrombin activity on vasopermeability and on coagulation especially when the major natural anticoagulant pathways are impaired.
authors.affiliation: Department of Internal Medicine, IRCCS Foundation Ospedale Maggiore Policlinico, University of Milan, Italy.
authors.forename: Sonia
authors.initials: S
authors.lastname: Caccia
authors.forename: Roberto
authors.initials: R
authors.lastname: Castelli
authors.forename: Diana
authors.initials: D
authors.lastname: Maiocchi
authors.forename: Luigi
authors.initials: L
authors.lastname: Bergamaschini
authors.forename: Massimo
authors.initials: M
authors.lastname: Cugno
mesh_terms: D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001777:Blood Coagulation / Q000187:drug effects* / Q000502:physiology; D002199:Capillary Permeability / Q000187:drug effects; D002478:Cells, Cultured; D050718:Complement C1 Inhibitor Protein / Q000378:metabolism / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006487:Hemostasis / Q000187:drug effects; D006493:Heparin / Q000378:metabolism / Q000494:pharmacology; D019763:Heparin Lyase / Q000378:metabolism / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D011485:Protein Binding / Q000187:drug effects; D019087:Selectins / Q000378:metabolism; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D000911:Antibodies, Monoclonal; D050718:Complement C1 Inhibitor Protein; D019087:Selectins; D006493:Heparin; D013917:Thrombin; D019763:Heparin Lyase