Correlation between trypsin binding to a specific receptor and prostacyclin production in cultured vascular endothelial cells.
The correlation between the binding and processing of trypsin and its effect on prostacyclin (PGI2) production in cultured adult bovine aortic endothelial (ABAE) cells was studied. ABAE cells demonstrated an ability to produce PGI2 in a dose-response manner to trypsin at the range of 0.1-2.0 micrograms/ml with a saturation at a concentration of 1 microgram/ml. Likewise, 125I-trypsin binding to the cultured cells increased in a dose-response way and reached saturation at a concentration of about 1 microgram/ml; 125I-trypsin was bound to a specific high-affinity cell-surface receptor with a dissociation constant (Kd) of 1.5 X 10(-8) M and each of the confluent ABAE cells has about 1.2 X 10(5) such receptors sites. The cell-surface receptor for trypsin displayed specific characteristics and an excess amount of unlabeled trypsin successfully abolished 125I-trypsin binding while thrombin in excess failed to compete for 125I-trypsin binding. Only a small fraction of the cell-surface-bound 125I-trypsin was internalized and subsequently degraded by ABAE cells as compared to the process of 125I-trypsin internalization by human skin fibroblasts (HSF). This study demonstrated that the stimulatory effect of trypsin on prostacyclin production and release by ABAE cells might be mediated by a specific cell-surface receptor for trypsin on these cells distinct from the thrombin receptor.
Savion
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Naveh-Floman
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D015121:6-Ketoprostaglandin F1 alpha / Q000032:analysis; D000818:Animals; D001011:Aorta / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D015232:Dinoprostone; D004727:Endothelium / Q000378:metabolism; D011464:Epoprostenol / Q000096:biosynthesis*; D007700:Kinetics; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D011458:Prostaglandins E / Q000032:analysis; D044464:Receptor, PAR-2; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014357:Trypsin / Q000378:metabolism* / Q000494:pharmacology
D011458:Prostaglandins E; D044464:Receptor, PAR-2; D011956:Receptors, Cell Surface; D015121:6-Ketoprostaglandin F1 alpha; D011464:Epoprostenol; D014357:Trypsin; D015232:Dinoprostone