Regulation of RAS in human platelets. Evidence that activation of RAS is not sufficient to lead to ERK1-2 phosphorylation.
In this study, we show that the G protein-coupled receptor agonist thrombin, the glycoprotein VI agonist convulxin, and the cytokine receptor Mpl agonist thrombopoietin (TPO) are able to induce activation of RAS in human platelets. Recruitment of GRB2 by tyrosine-phosphorylated proteins in response to TPO and convulxin but not by thrombin occurred with a similar time-course to RAS activation, consistent with a causal relationship. On the other hand, activation of ERK2 by thrombin and convulxin is delayed and also inhibited by the protein kinase C inhibitor Ro-31 8220, whereas RAS activation is unaffected. Further evidence for differential regulation of RAS and ERK is provided by the observations that TPO, which activates RAS but not protein kinase C, does not activate ERK, and that the inhibitor of SRC kinases PP1 inhibits activation of RAS but not ERK2 in response to thrombin. Our results demonstrate that activation of RAS is not necessarily coupled to ERK in human platelets.
Tulasne
David
D
Department of Pharmacology, University of Oxford, UK. david.tuslane@pharmacology.oxford.ac.uk
Bori
Teresa
T
Watson
Steve P
SP
D048868:Adaptor Proteins, Signal Transducing; D001792:Blood Platelets / Q000378:metabolism*; D003435:Crotalid Venoms / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D051380:GRB2 Adaptor Protein; D006801:Humans; D037181:Lectins, C-Type; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D010766:Phosphorylation; D010974:Platelet Aggregation; D011506:Proteins / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology; D018631:ras Proteins / Q000378:metabolism*; D019061:src-Family Kinases / Q000502:physiology
D048868:Adaptor Proteins, Signal Transducing; D003435:Crotalid Venoms; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D037181:Lectins, C-Type; D011506:Proteins; C028890:convulxin; D013926:Thrombopoietin; D019061:src-Family Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases; D013917:Thrombin; D018631:ras Proteins