title: Evidence for two distinct G-protein-coupled ADP receptors mediating platelet activation.
abstract: The identity of the receptors mediating platelet activation by ADP remains elusive. To distinguish between platelet ADP receptor subtypes, the effects of antagonists on platelet responses and the cloned P2Y1 receptor, a putative platelet ADP receptor, have been investigated. 2-methylthio-AMP (2MeSAMP), an inhibitor of ADP-dependent platelet aggregation, antagonized ADP-mediated inhibition of adenylyl cyclase, competed with binding of [3H]2-methylthio-ADP and inhibited the stimulation of [35S]GTPgammaS binding. 2MeSAMP did not inhibit platelet shape change and was only a weak antagonist of intracellular calcium mobilization in platelets or in cells expressing the cloned human P2YI receptor. By contrast, the P2Y1 receptor antagonist adenosine 3',5'-diphosphate (A3P5P) inhibited ADP-induced platelet aggregation, completely abolished shape change, but did not antagonize ADP effects on cyclic AMP generation or [3H]2-methylthio-ADP binding. However, A3P5P antagonized intracellular calcium mobilization in platelets and cells expressing the cloned P2Y1 receptor. Furthermore, using a specific monoclonal antibody and flow cytometry, P2Y1 receptor protein was detected on human platelets. These results support the existence of two G protein-coupled ADP receptors mediating platelet aggregation, one of which is coupled to Gi proteins and blocked by 2MeSAMP, whereas the second receptor is similar or identical to P2Y1 and coupled to Gq.
authors.affiliation: COR Therapeutics, Inc., South San Francisco, CA 94080, USA. mjantzen@corr.com
authors.forename: H M
authors.initials: HM
authors.lastname: Jantzen
authors.forename: L
authors.initials: L
authors.lastname: Gousset
authors.forename: V
authors.initials: V
authors.lastname: Bhaskar
authors.forename: D
authors.initials: D
authors.lastname: Vincent
authors.forename: A
authors.initials: A
authors.lastname: Tai
authors.forename: E E
authors.initials: EE
authors.lastname: Reynolds
authors.forename: P B
authors.initials: PB
authors.lastname: Conley
mesh_terms: D001792:Blood Platelets / Q000502:physiology*; D005434:Flow Cytometry; D019204:GTP-Binding Proteins / Q000502:physiology; D006801:Humans; D015539:Platelet Activation; D018048:Receptors, Purinergic P2 / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D018048:Receptors, Purinergic P2; D019204:GTP-Binding Proteins