Involvement of Na+/Ca2+ exchanger in inside-out signaling through the platelet integrin IIbbeta3.
The platelet integrin IIbbeta3 has become a new target for the treatment of pathological thrombosis. It becomes apparent that the affinity of IIbbeta3 for its ligands is dynamically regulated by inside-out signaling. However, the components that couple diverse intracellular signals to the cytoplasmic domains of IIbbeta3 remain obscure. Employing a chymotrypsin-induced IIbbeta3 activation model, we previously proposed the hypothesis that Na+/Ca2 + exchanger (NCX) may be involved in inside-out signaling (Shiraga et al: Blood 88:2594, 1996). In the present study, employing two unrelated Na+/Ca2+ exchange inhibitors, 3',4'-dichlorobenzamil (DCB) and bepridil, we investigated the role of NCX in platelet activation induced by various agonists in detail. Both inhibitors abolished platelet aggregation induced by all agonists examined via the inhibition of IIbbeta3 activation. Moreover, these inhibitors abolished IIbbeta3 activation induced by phorbol 12-myristate 13-acetate or A23187. On the other hand, neither of these inhibitors showed apparent inhibitory effects on protein phosphorylation of pleckstrin or myosin light chain, or an increase in intracellular calcium ion concentrations evoked by 0.1 U/mL thrombin. These effects of the NCX inhibitors are in striking contrast to those of protein kinase C inhibitor, Ro31-8220. Biochemical and ultrastructural analyses showed that NCX inhibitors, particularly DCB, made platelets "thrombasthenic". These findings suggest that the NCX is involved in the common steps of inside-out signaling through integrin IIbbeta3.
Shiraga
M
M
Second Department of Internal Medicine, Osaka University Medical School and Department of Blood Transfusion, Osaka University Hospital, Osaka, Japan, USA.
Tomiyama
Y
Y
Honda
S
S
Suzuki
H
H
Kosugi
S
S
Tadokoro
S
S
Kanakura
Y
Y
Tanoue
K
K
Kurata
Y
Y
Matsuzawa
Y
Y
D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D015764:Bepridil / Q000494:pharmacology; D001792:Blood Platelets / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002918:Chymotrypsin / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D017136:Ion Transport / Q000187:drug effects; D018994:Myosin Light Chains / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism; D010750:Phosphoproteins; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000502:physiology*; D011499:Protein Processing, Post-Translational; D012701:Serotonin / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D012964:Sodium / Q000378:metabolism*; D019831:Sodium-Calcium Exchanger / Q000037:antagonists & inhibitors / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D001798:Blood Proteins; D018994:Myosin Light Chains; D019007:P-Selectin; D010750:Phosphoproteins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019831:Sodium-Calcium Exchanger; C039077:platelet protein P47; C041470:3',4'-dichlorobenzamil; D012701:Serotonin; D000001:Calcimycin; D015764:Bepridil; D000584:Amiloride; D005340:Fibrinogen; D012964:Sodium; D002918:Chymotrypsin; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium