Identification of small GTP-binding rab proteins in human platelets: thrombin-induced phosphorylation of rab3B, rab6, and rab8 proteins.
The activation of platelets by specific agonists is a tightly regulated mechanism that leads to the secretion of the dense- and alpha-granule contents. Platelets have been shown to possess small GTP-binding proteins thought to be involved in central biological processes; however, no rab proteins, which may regulate the exocytic process at different stages, have been reported. This study has shown that rab1, rab3B, rab4, rab6, and rab8 proteins, but not rab3A protein, were present in platelets and in endothelial cells. To probe their functional significance in platelets, rab3B, rab6, and rab8 proteins were further characterized with regard to their intracellular localization and their phosphorylation properties. Whereas rab3B protein was found to be mainly cytosolic, rab6 and rab8 proteins were preferentially targeted to the plasma membrane and to the alpha granules. The activation of platelets by thrombin, a potent inducer of secretion, resulted in the phosphorylation of rab3B, rab6, and rab8 proteins, whereas no phosphorylation was observed in the presence of prostaglandin E1, which stimulates cAMP-dependent protein kinase and inhibits the secretion process. These findings provide evidence that members of the subfamily of rab proteins, rab6 and rab8, are localized in platelets to one type of specific secretory vesicle, the alpha granule, and would suggest their possible implication in the secretion process through phosphorylation mechanisms.
Karniguian
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Institut National de la Santé et de la Recherche Médicale, U 248, Faculté de Médecine, Paris, France.
Zahraoui
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Tavitian
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D000595:Amino Acid Sequence; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000378:metabolism / Q000648:ultrastructure; D004730:Endothelium, Vascular / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D010766:Phosphorylation; D015539:Platelet Activation; D013347:Subcellular Fractions / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology*; D014471:Umbilical Veins; D020691:rab GTP-Binding Proteins; D020692:rab3 GTP-Binding Proteins
D013917:Thrombin; D019204:GTP-Binding Proteins; C082377:RAB8A protein, human; D020691:rab GTP-Binding Proteins; D020692:rab3 GTP-Binding Proteins