Total inhibition of phospholipase C and phosphatidylinositol 3-kinase by okadaic acid in thrombin-stimulated platelets.
The strong inhibition of thrombin-induced platelet functions induced by okadaic acid is not correlated with the partial modification of pleckstrin phosphorylation, which remains still phosphorylated two min after stimulation, indicating that protein kinase C is not affected by okadaic acid. We then investigated the effect of okadaic acid on platelet lipid metabolism. Our data indicate that inhibition indeed strongly affects phosphatidic acid as well as phosphatidylinositol 3,4-bisphosphate synthesis at low concentrations of okadaic acid, and phosphatidylinositol 4,5-bisphosphate at higher concentrations. Since thrombin-induced tyrosine phosphorylations were completely inhibited in the presence of okadaic acid, as a consequence, phosphatidylinositol 3-kinase was no longer detected in antiphosphotyrosine immunoprecipitates, thus explaining the absence of phosphatidylinositol, 3,4-bisphosphate synthesis. Finally, okadaic acid inhibited thrombin-induced fibrinogen binding, indicating that serine/threonine phosphatases may affect the inside-out signalling which regulates the alpha 11bb3 integrin, downstream protein kinase C activation.
Mauco
G
G
INSERM Unité 326, Hôpital Purpan, Toulouse, France.
Artçanuthurry
V
V
Pidard
D
D
Grelac
F
F
Maclouf
J
J
Lévy-Tolédano
S
S
D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D008934:Mitogens / Q000494:pharmacology*; D019319:Okadaic Acid / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010750:Phosphoproteins; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors*; D019000:Phosphotyrosine / Q000378:metabolism; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D011233:Precipitin Tests; D013917:Thrombin / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors*; D014443:Tyrosine / Q000378:metabolism
D001798:Blood Proteins; D004791:Enzyme Inhibitors; D008934:Mitogens; D018129:Phosphatidylinositol Phosphates; D010750:Phosphoproteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; C039077:platelet protein P47; D019319:Okadaic Acid; D019000:Phosphotyrosine; D014443:Tyrosine; D005340:Fibrinogen; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D010738:Type C Phospholipases; D013917:Thrombin