ACTIVE-SITE-DEPENDENT, THROMBIN-INDUCED RELEASE OF ADENINE NUCLEOTIDES FROM CULTURED HUMAN ENDOTHELIAL CELLS*

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Abstract
The effect of thrombin on release of adenine nucleotides is studied in cultured cell monolayers from the human umbilical vein. Thrombin-induced release of radioactivity from endothelial cells is dose-dependent and saturable with maximal response seen at 1 x 10(-8) M thrombin. The products are identified by thin-layer chromatography as adenine nucleotides. Diisopropylphosphoryl thrombin, which is enzymatically inactive, does not cause release of tritium. A 50-fold excess of diisopropylphosphoryl-thrombin, despite causing 98 percent inhibition of binding of 125I-thrombin to its high-affinity binding sites, does not inhibit thrombin-induced release. We conclude that (1) thrombin causes release from endothelial cells of adenine nucleotides and that (2) high-affinity, active-site-independent binding of thrombin is not involved in this process.