Adhesion receptor activation of phosphatidylinositol 3-kinase. von Willebrand factor stimulates the cytoskeletal association and activation of phosphatidylinositol 3-kinase and pp60c-src in human platelets.
The cytoskeleton participates in the coordinated regulation of intracellular signaling molecules, following agonist stimulation of cells. We have demonstrated that von Willebrand factor (vWF) induced the cytoskeletal association and activation of phosphatidylinositol 3-kinase (PtdIns 3-kinase) in human platelets. The activation of PtdIns 3-kinase coincided with the tyrosine phosphorylation of multiple platelet proteins, as assessed by anti-phosphotyrosine immunoblotting. One of these tyrosine-phosphorylated proteins, pp60c-src, became specifically enriched in the cytoskeletal fraction of vWF-stimulated platelets. The vWF-stimulated cytoskeletal association of PtdIns 3-kinase and pp60c-src required platelet stirring and aggregation, was specifically blocked by an anti-GPIb monoclonal antibody, and was not observed in platelets lacking the glycoprotein Ib/IX complex (Bernard-Soulier syndrome). Pretreatment of normal platelets with 5 mM EDTA (37 degrees C for 90 min) or RGDS (2 mM), which disrupts the binding of various adhesive proteins to platelet integrins and inhibits fibrinogen-mediated platelet aggregation, did not alter the vWF-stimulated activation and cytoskeletal association of PtdIns 3-kinase and pp60c-src. Pretreatment of platelets with acetylsalicylic acid (1 mM) completely abolished vWF-stimulated production of thromboxane A2, dense granule release, and the activation of protein kinase C, without altering the activation and cytoskeletal translocation of PtdIns 3-kinase and pp60c-src. Our results suggest that vWF binding to the platelet adhesion receptor glycoprotein Ib/IX can mediate activation and translocation of both tyrosine and lipid kinase(s) independent of other agonists.
Jackson
S P
SP
Department of Medicine, Monash Medical School, Box Hill Hospital, Victoria, Australia.
Schoenwaelder
S M
SM
Yuan
Y
Y
Rabinowitz
I
I
Salem
H H
HH
Mitchell
C A
CA
D001241:Aspirin / Q000494:pharmacology; D001692:Biological Transport; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology*; D002451:Cell Compartmentation; D003599:Cytoskeleton / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D004789:Enzyme Activation; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D019869:Phosphatidylinositol 3-Kinases; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism*; D010974:Platelet Aggregation / Q000502:physiology; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000378:metabolism; D016024:Receptors, Cytoadhesin / Q000378:metabolism*; D012310:Ristocetin / Q000494:pharmacology; D015398:Signal Transduction; D014841:von Willebrand Factor / Q000494:pharmacology
D017261:Glycosylphosphatidylinositols; D010980:Platelet Membrane Glycoproteins; D016024:Receptors, Cytoadhesin; D014841:von Willebrand Factor; D012310:Ristocetin; D004492:Edetic Acid; D019869:Phosphatidylinositol 3-Kinases; D017853:Phosphotransferases (Alcohol Group Acceptor); D016392:Proto-Oncogene Proteins pp60(c-src); D001241:Aspirin