Tyrosine phosphorylation and Syk activation are involved in thrombin-induced aggregation of epinephrine-potentiated platelets.
Thrombin and epinephrine in combination exert synergistic effects on platelet activation. On the other hand, tyrosine phosphorylation and activation of tyrosine kinases including Syk have been shown to play a critical role in the induction of platelet responses to thrombin stimulation. This study investigated the role of tyrosine phosphorylation and Syk activation in the synergistic mechanisms between thrombin and epinephrine. Although epinephrine alone (4 microM) slightly induced protein-tyrosine phosphorylation and Syk activation, the presence of epinephrine caused a shift to the left in the dose-dependence of thrombin (0.01-0.5 U/ml)-induced tyrosine phosphorylation and Syk activation, as well as platelet aggregation. Phenoxybenzamine, an alpha-adrenoceptor antagonist, canceled this potentiation by epinephrine. Since platelets dominantly express alpha 2-adrenoceptor, this result indicates that epinephrine acts through the occupancy of alpha 2-adrenoceptor. Furthermore, pretreatment with a tyrosine kinase inhibitor, genistein, or a cAMP-elevating agent, prostacyclin (PGI2), significantly reduced these synergistic effects of epinephrine. Taken together, our results suggested that the potentiation by epinephrine may be mediated via enhancement of tyrosine phosphorylation and Syk activation, in part through a decrease of intracellular cAMP levels.
Wang
X
X
Department of Biochemistry, Kobe University, School of Medicine.
Yanagi
S
S
Yang
C
C
Inatome
R
R
Yamamura
H
H
D000316:Adrenergic alpha-Agonists / Q000494:pharmacology*; D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects* / Q000201:enzymology; D004357:Drug Synergism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004792:Enzyme Precursors / Q000378:metabolism*; D004837:Epinephrine / Q000494:pharmacology*; D011464:Epoprostenol / Q000494:pharmacology; D019833:Genistein; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007529:Isoflavones / Q000494:pharmacology; D010643:Phenoxybenzamine / Q000494:pharmacology; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000072377:Syk Kinase; D013917:Thrombin / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*
D000316:Adrenergic alpha-Agonists; D000317:Adrenergic alpha-Antagonists; D004791:Enzyme Inhibitors; D004792:Enzyme Precursors; D047908:Intracellular Signaling Peptides and Proteins; D007529:Isoflavones; D010975:Platelet Aggregation Inhibitors; D010643:Phenoxybenzamine; D014443:Tyrosine; D011464:Epoprostenol; D019833:Genistein; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; D013917:Thrombin; D004837:Epinephrine