Receptor-mediated regulation of leukotriene C4 synthase activity in human platelets.
Human platelets possess a specific membrane-bound leukotriene (LT) C4 synthase, which catalyzes the conversion of LTA4 to LTC4. Stimulation of the receptors for thrombin, collagen or thromboxane A2 provoked inhibition of this enzyme, as judged by suppressed transformation of exogenous LTA4 to LTC4. Similarly, direct activation of protein kinase (PK) C with nanomolar concentrations of 4 beta-phorbol 12-myristate 13-acetate (PMA) inhibited the production of LTC4. Kinetic studies demonstrated that the inhibition induced by thrombin and PMA was non-competitive. Elevation of intracellular cAMP levels with carbacyclin did not affect basal LTC4 formation, but abolished the attenuation of platelet LTC4 synthase activity induced by the thromboxane receptor agonist U-46619. The unselective protein kinase inhibitor staurosporine prevented both receptor-mediated and PMA-induced suppression of LTC4 formation. In contrast, two selective PKC inhibitors, Ro 31-8220 and GF 109203X, reversed the inhibitory effect provoked by PMA, but failed to prevent thrombin-induced inhibition. Furthermore, the protein tyrosine phosphatase inhibitor, sodium orthovanadate, induced dose-dependent inhibition of LTC4 production in platelet sonicates. In conclusion, receptor-mediated activation of human platelets leads to decreased LTC4 synthase activity via phosphoregulation. Although the present results demonstrate that platelet LTC4 synthase can be regulated via PKC-dependent events, alternative mechanisms appears to be involved in the physiological regulation of this enzyme. The findings suggest the possible importance of protein tyrosine phosphorylations in this process.
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
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Tornhamre
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Edenius
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Lindgren
D001792:Blood Platelets / Q000201:enzymology*; D011464:Epoprostenol / Q000031:analogs & derivatives / Q000494:pharmacology; D005982:Glutathione Transferase / Q000378:metabolism*; D006801:Humans; D017572:Leukotriene A4 / Q000378:metabolism; D017997:Leukotriene C4 / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000502:physiology; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology
D017572:Leukotriene A4; D017997:Leukotriene C4; C026678:carboprostacyclin; D011464:Epoprostenol; D005982:Glutathione Transferase; D011493:Protein Kinase C; D013917:Thrombin; C041116:leukotriene-C4 synthase; D013755:Tetradecanoylphorbol Acetate