Related Experiment Videos
Thrombin-induced Ca2+ influx and protein tyrosine phosphorylation in endothelial cells is inhibited by herbimycin A
H J Kruse1, E V Negrescu, P C Weber
1Institut für Prophylaxe und Epidemiologie der Kreislaufkrankheiten, University of Munich, Germany.
During endothelial cell activation, alpha-thrombin elicits both Ca2+ release from internal stores and influx of external Ca2+ across the plasma membrane. The mechanisms of alpha-thrombin-induced Ca2+ entry into endothelial cells are unclear. Therefore, effects of the specific tyrosine kinase inhibitor herbimycin A on protein tyrosine phosphorylation and on intracellular Ca2+ transients were studied in alpha-thrombin-stimulated human umbilical vein endothelial cells. alpha-Thrombin caused significant tyrosine phosphorylation of mainly two proteins and evoked typical biphasic changes of free cytosolic Ca2+ concentration. We show that 24 h pretreatment with herbimycin A inhibited alpha-thrombin-induced endothelial protein tyrosine phosphorylation. Moreover, herbimycin A significantly attenuated alpha-thrombin-induced Ca2+ influx but not release from internal stores. The data suggest that protein tyrosine phosphorylation by alpha-thrombin is involved in the regulation of alpha-thrombin-induced Ca2+ influx into endothelial cells.
During endothelial cell activation, alpha-thrombin elicits both Ca2+ release from internal stores and influx of external Ca2+ across the plasma membrane. The mechanisms of alpha-thrombin-induced Ca2+ entry into endothelial cells are unclear. Therefore, effects of the specific tyrosine kinase inhibitor herbimycin A on protein tyrosine phosphorylation and on intracellular Ca2+ transients were studied in alpha-thrombin-stimulated human umbilical vein endothelial cells. alpha-Thrombin caused significant tyrosine phosphorylation of mainly two proteins and evoked typical biphasic changes of free cytosolic Ca2+ concentration. We show that 24 h pretreatment with herbimycin A inhibited alpha-thrombin-induced endothelial protein tyrosine phosphorylation. Moreover, herbimycin A significantly attenuated alpha-thrombin-induced Ca2+ influx but not release from internal stores. The data suggest that protein tyrosine phosphorylation by alpha-thrombin is involved in the regulation of alpha-thrombin-induced Ca2+ influx into endothelial cells.