Signaling by ephrinB1 and Eph kinases in platelets promotes Rap1 activation, platelet adhesion, and aggregation via effector pathways that do not require phosphorylation of ephrinB1.
We have previously shown that platelets express 2 receptor tyrosine kinases, EphA4 and EphB1, and the Eph kinase ligand, ephrinB1, and proposed that transcellular Eph/ephrin interactions made possible by the onset of platelet aggregation promote the further growth and stability of the hemostatic plug. The present study examines how this might occur. The results show that clustering of either ephrinB1 or EphA4 causes platelets to adhere to immobilized fibrinogen via alpha(IIb)beta(3). Adhesion occurs more slowly than with adenosine diphosphate (ADP) and requires phosphatidylinositol 3 (PI3)-kinase and protein kinase C activity but not ephrinB1 phosphorylation. By itself, Eph and ephrin signaling is insufficient to cause aggregation or the binding of soluble fibrinogen, but it can potentiate aggregation initiated by a Ca(++) ionophore or by agonists for thrombin and thromboxane receptors. It also enhances Rap1 activation without requiring ADP secretion, ephrinB1 phosphorylation, or the activation of PI3-kinase and Src. From this we conclude that (1) Eph/ephrin signaling enhances the ability of platelet agonists to cause aggregation provided that those agonists can increase cytosolic Ca(++); (2) this is accomplished in part by activating Rap1; and (3) these effects require oligomerization of ephrinB1 but not phosphotyrosine-based interactions with the ephrinB1 cytoplasmic domain.
Department of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Nicolas
N
Prévost
Donna S
DS
Woulfe
Massimiliano
M
Tognolini
Takako
T
Tanaka
Wenying
W
Jian
Ryan R
RR
Fortna
Hong
H
Jiang
Lawrence F
LF
Brass
D001792:Blood Platelets / Q000378:metabolism*; D036385:Ephrin-A4 / Q000235:genetics / Q000378:metabolism; D036387:Ephrin-B1 / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D010973:Platelet Adhesiveness / Q000502:physiology; D010974:Platelet Aggregation / Q000502:physiology*; D011494:Protein Kinases / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism*
D036385:Ephrin-A4; D036387:Ephrin-B1; D011993:Recombinant Fusion Proteins; D011494:Protein Kinases; D020827:rap1 GTP-Binding Proteins