Differential involvement of membrane (Na-K)ATPase in thrombin- and trypsin-mediated platelet activation.
The possibility that platelet activation may also involve membrane (Na-K)ATPase was investigated by testing the effects of both proteinases on platelet shape change and aggregation in the absence and presence of the specific (Na-K)ATPase inhibitor ouabain. Ouabain (8 to 80 microM) completely antagonized trypsin-induced platelet shape change and aggregation when it was preincubated with platelet suspension before the addition of trypsin. Unlike trypsin, thrombin-induced platelet activation was significantly enhanced by ouabain. It was also observed that on partially purified beef heart (Na-K)ATPase preparation, thrombin significantly enhanced the enzyme inhibition caused by submaximal inhibitory concentrations of ouabain. Soybean trypsin inhibitor (4 micrograms/ml) employed as the agent capable to counteract proteinase effects on the (Na-K)ATPase, was shown both to prevent and antagonize the platelet activation induced by trypsin (0.3 to 1.5 micrograms/ml), but it failed to modify the responses evoked by thrombin. It is concluded that membrane (Na-K)ATPase is involved differently in platelet activation by trypsin and thrombin probably because receptor sites to which either proteinase on the platelet surface binds, are distinct. Direct enzyme involvement is indeed apparent only in trypsin-induced platelet activation.
Finotti
P
P
Department of Pharmacology, University of Padova, Italy.
D001665:Binding Sites; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D010042:Ouabain / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects*; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors*; D013917:Thrombin / Q000494:pharmacology*; D014357:Trypsin / Q000494:pharmacology*; D014361:Trypsin Inhibitors / Q000494:pharmacology
D010975:Platelet Aggregation Inhibitors; D014361:Trypsin Inhibitors; D010042:Ouabain; D014357:Trypsin; D013917:Thrombin; D000254:Sodium-Potassium-Exchanging ATPase