Reversal of thrombin-induced myosin phosphorylation and the assembly of cytoskeletal structures in platelets by the adenylate cyclase stimulants prostaglandin D2 and forskolin.
Stimulation of platelets by thrombin causes an increase in the amount of cytoskeleton proteins insoluble in 1% Triton X-100, i.e. myosin, actin, actin-binding protein, an alpha-actinin-like protein of Mr = 105,000, unidentified polypeptides of Mr = 150,000, 31,00, and under some conditions, 56,000. Concurrently the Mr = 20,000 light chains of myosin and a cytoplasmic Mr = 42,000 polypeptide are phosphorylated, presumably by calmodulin-Ca2+-dependent myosin light chain kinase and a phospholipid-Ca2+-dependent kinase, respectively. The adenylate cyclase stimulators prostaglandin D2 (PGD2) and forskolin increased platelet cyclic AMP and prevented the phosphorylation of these polypeptides and the increase in Triton-insoluble cytoskeleton proteins. When added to platelets after stimulation by thrombin they caused rapid complete reversal of myosin light chain and Mr = 42,000 polypeptide phosphorylation; simultaneously the association of myosin with the cytoskeleton proteins and the increase in the content of each of the Triton-insoluble cytoskeleton proteins (except the Mr = 56,000 polypeptide) was reversed. The amount of Triton-insoluble myosin was affected more readily by PGD2 or forskolin than were the other proteins. Increasing thrombin from 0.1 to 1.0 unit/ml inhibited all the responses to PGD2 and forskolin possibly due to concentration-dependent effects of thrombin that inhibit adenylate cyclase. These results suggest that cytoskeleton assembly and activation of the contractile apparatus in intact platelets are readily reversible by cyclic AMP-dependent reactions.
D000262:Adenylyl Cyclases / Q000097:blood; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D001798:Blood Proteins / Q000378:metabolism; D005576:Colforsin; D004224:Diterpenes / Q000494:pharmacology*; D006801:Humans; D009218:Myosins / Q000378:metabolism*; D010766:Phosphorylation; D015230:Prostaglandin D2; D011453:Prostaglandins / Q000494:pharmacology*; D011457:Prostaglandins D / Q000494:pharmacology*; D013806:Theophylline / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology*
D001798:Blood Proteins; D004224:Diterpenes; D011453:Prostaglandins; D011457:Prostaglandins D; D005576:Colforsin; D013806:Theophylline; D013917:Thrombin; D009218:Myosins; D000262:Adenylyl Cyclases; D015230:Prostaglandin D2