Phosphatidylinositol(3, 4, 5) -Trisphosphate Stimulates Phosphorylation of Pleckstrin in Human Platelets (*)

We have reported that platelets exposed to thrombin or thrombin receptor-directed ligand activate phospholipase C and rapidly accumulate phosphatidylinositol(3, 4, 5) -trisphosphate (PtdIns(3,4,5)P) and phosphatidylinositol (3, 4) -bisphosphate (PtdIns(3,4)P) as a function of the activation of phosphoinositide (PI) 3-kinases in a GTP-binding protein-dependent manner. In such platelets, serine- and threonine-directed phosphorylation of pleckstrin also occurs and has been attributed to protein kinase C activation. We now report that the phosphorylation of pleckstrin is partially dependent upon PI 3-kinase. Pleckstrin phosphorylation in response to thrombin receptor stimulation is progressively susceptible to inhibition by wortmannin, a potent and specific inhibitor of platelet PI 3-kinases. PI 3-kinase thus seems to play a gradually increasing role in promoting pleckstrin phosphorylation. The IC for wortmannin in inhibiting SFLLRN-stimulated 3-phosphorylated phosphoinositide accumulation is 10 nM, and that (i.e. 50% of maximum inhibition) for inhibiting pleckstrin phosphorylation is 15 nM. Synthetic PtdIns(3,4,5)P, when added to saponin-permeabilized (but not intact) platelets, causes wortmannin-insensitive phosphorylation of pleckstrin. PtdIns(3,4,5)P also overcomes the inhibition by wortmannin of thrombin- or guanosine 5′-3-O-(thio)trisphosphate-stimulated pleckstrin phosphorylation. In contrast, PtdIns(4,5)P or inositol (1, 3, 4, 5) -tetrakisphosphate are ineffective in these respects. The pattern of phosphorylation of pleckstrin activated by PtdIns(3,4,5)P is not distinguishable from that of pleckstrin phosphorylated in intact platelets exposed to protein kinase C-activating β-phorbol myristate acetate, mimicking diacylglycerol. Activation of protein kinase(s) by PtdIns(3,4,5)P thus offers a route for pleckstrin phosphorylation in vivo that is an alternative to activation of phospholipase C diacylglycerol protein kinase C.

• Publication year

  1995

• Venue

  Journal of Biological Chemistry

• Publication date

  1995-09-29

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1074/JBC.270.39.22807PMID 7559410
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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