The cleaved peptide of PAR1 is a more potent stimulant of platelet-endothelial cell adhesion than is thrombin
Department of Pediatrics; Department of Surgery
Medical Subject Headings
Cell Adhesion; Cell Division; Endothelium; Hemostatics; Humans; Peptide Fragments; Platelet Adhesiveness; Receptor, PAR-1; Receptors, Thrombin; Saphenous Vein; Thrombin; Venous Thrombosis
Hematology | Oncology | Pediatrics
PURPOSE: Platelet-endothelial cell adhesion is an important pathologic response to vessel injury or inflammation. On binding to its endothelial or platelet G protein-linked seven-transmembrane domain receptor, protease-activated receptor-1 (PAR1), thrombin releases a 41-amino acid peptide (TR(1-41)). We examined the effect of TR(1-41) on platelet activation and on platelet-endothelial cell adhesion.
METHODS: A monolayer of confluent human saphenous vein endothelial cells was incubated with washed human platelets. Platelets were stimulated with either TR(1-41), TR(21-41), scrambled TR(1-41), adenosine diphosphate (ADP)-epinephrine (EPI), thrombin, or thrombin receptor activating peptide (TRAP). Platelet activation was identified with flow cytometry. The magnitude of platelet-endothelial cell adhesion was determined with a laser scanning cytometer that scanned the monolayer of endothelial cells and identified fluorescently bound platelets. RESULTS: Maximal thrombin stimulation (0.1 U/mL) induced a threefold increase in platelets bound to endothelial cells compared with buffer alone. Stimulation with TR(1-41) (20 mmol/L) tripled the number of platelets bound to endothelial cells compared with thrombin. Scrambled sequence of TR(1-41) (20 mmol/L) and TR(21-41) (20 mmol/L), neither of which induces platelet activation, had minimal effect on platelet adhesion. Both TRAP (20 mmol/L) and ADP-EPI (20 mmol/L) induced less platelet-endothelial cell adhesion than did thrombin. TR(1-41)-induced platelet-endothelial cell adhesion was partially blocked by glycoprotein (GP)IIb-IIIa-specific monoclonal antibody, 10E5 (10 mg/mL).
CONCLUSIONS: TR(1-41), the cleaved peptide of PAR1, is a more potent stimulant of platelet-endothelial cell adhesion than is thrombin, TRAP, or ADP-EPI, and this adhesion is at least in part mediated by the platelet GPIIb-IIIa receptor.
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Citation: J Vasc Surg. 2003 Feb;37(2):440-5. doi 10.1067/mva.2003.129
Claytor, R. Brannon; Michelson, Alan D.; Li, Jian-Ming; Frelinger, Andrew L. III; Rohrer, Michael J.; Garnette, Charles S. C.; Barnard, Marc R.; Krueger, Lori A.; and Furman, Mark I., "The cleaved peptide of PAR1 is a more potent stimulant of platelet-endothelial cell adhesion than is thrombin" (2003). Hematology/Oncology. 44.