P2y(12), a new platelet ADP receptor, target of clopidogrel

• Published in: Biochemical and biophysical research communications Vol. 283; no. 2; p. 379
• Main Authors: Savi, P, Labouret, C, Delesque, N, Guette, F, Lupker, J, Herbert, J M
• Format: Journal Article
• Language: English
• Published: United States 04.05.2001
• Subjects: Adenosine Diphosphate - analogs & derivatives; Adenosine Diphosphate - metabolism; Adenylyl Cyclases - metabolism; Animals; Blood Platelets - drug effects; Blood Platelets - metabolism; CHO Cells; Cricetinae; Down-Regulation; Humans; Kinetics; Receptors, Purinergic P2 - blood; Receptors, Purinergic P2 - drug effects; Receptors, Purinergic P2 - genetics; Ticlopidine - analogs & derivatives; Ticlopidine - pharmacology; Transfection
• DOI: 10.1006/bbrc.2001.4816
• ISSN: 0006-291X

The binding characteristics of (33)P-2MeS-ADP, a stable analogue of ADP, were determined on CHO cells transfected with the human P2Y(12) receptor, a novel purinergic receptor. These transfected CHO cells displayed a strong affinity for (33)P-2MeS-ADP, the binding characteristics of which corresponded in all points to those observed on platelets. In particular, this receptor recognised purines with the following order of potency: 2MeS-ADP = 2MeS-ATP > ADP = ATPgammaS = ATP >> UTP, a binding profile which is similar to that obtained in platelets. The binding of (33)P-2MeS-ADP was antagonised by pCMPS but not by MRS2179 and FSBA, antagonists of P2Y(1) and aggregin, respectively. Moreover, the binding of (33)P-2MeS-ADP to these cells was strongly and irreversibly inhibited by the active metabolite of clopidogrel with a potency which was consistent with that observed for this compound on platelets. Like in platelets, 2MeS-ADP induced adenylyl cyclase down-regulation in these P2Y(12) transfected CHO cells, an effect which was absent in the corresponding non-transfected cells. As already shown in platelets, the active metabolite of clopidogrel antagonised 2MeS-ADP-induced inhibition of adenylyl cyclase on transfected cells. Our results confirm that P2Y(12) is the previously called "platelet P2t(AC)" receptor and show that this receptor is antagonised by the active metabolite of clopidogrel.