calcimycin and amastatin

The present study was undertaken to define clearly the receptor, which is responsible for the thrombin induced vWf release from HUVEC. Vu et al. reported that cleavage of the platelet thrombin receptor by thrombin resulted in a new N-terminus (SFLLRN...) which acts as tethered ligand (4). The free peptide activates platelets and induces rises in both cytosolic free Ca2+ and PGI2 production in HUVEC (10). HUVEC were incubated with thrombin, SFLLRN or other relevant substances. After incubation, the intracellular vWf content was compared with the vWf concentration in the supernatant. The intracellular vWf concentration was measured by microscope fluorometry and the concentrations in the supernatant with an ELISA. The thrombin stimulated cells showed 53% vWf antigen compared with control cells (100%). This result was well correlated with the 2.4 fold higher vWf concentrations measured in the supernatant of thrombin stimulated cells than in control cells. Also the addition of SFLLRN (1-60 microM) or trypsin (1-50 nM) increased vWf release from HUVEC in a dose dependent manner. These results indicate that thrombin induced vWf release from HUVEC is mediated through the activation of the tethered ligand receptor.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Calcimycin; Calcium; Cells, Cultured; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Humans; Ligands; Molecular Sequence Data; Oligopeptides; Peptide Fragments; Peptides; Protein Binding; Receptors, Thrombin; Thrombin; Trypsin; Umbilical Veins; von Willebrand Factor