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Thrombin-induced changes in PA, monophosphoinositide (MPI) , diphosphoinositide (DPI) and triphosphoinositide (TPI) of washed rabbit platelets have been examined. Platelets prelabeled by incubation with 32P-orthophosphate, 3H-glycerol, 3H-inositol or 14C-arachidonate were exposed to 0.33 u/ml thrombin for one minute, and the phospholipids extracted and separated by thin layer chromatography. Measurement of absolute amounts of PA and MPI by phosphorus assay showed that PA increased by 180% while MPI decreased by 15%.  The changes in MPI with 3H-G, 3H-I and 14C-AA and in 1,2-diacyl glycerol (DG) indicate that some MPI may be converted to PA via DG. Changes in the 3H-G and 3H-I labeling of DPI and TPI suggest that the changes observed with 32P-Iabeled platelets are a result of the turnover of the phosphorylinositol moiety. The increase in AA with 14C-AA indicates that some of the decrease in MPI may be due to the formation of lyso MPI and free fatty acid. These results indicate that thrombin stimulation of platelets may affect inositol phospholipid metabolism through three pathways: (1) involving PA, DG and MPI; (2) the cleavage of free fatty acids from MPI; (3) turnover of the ester phosphates on DPI and TPI.

Thrombin-Induced Changes in Phosphatidic ACID (PA) and Phosphoinositides