title: Labeling of A2A adenosine receptors in human platelets by use of the new nonxanthine antagonist radioligand [3H]SCH 58261.
abstract: The present study describes the binding to human platelet A2A adenosine receptors of the new potent and selective antagonist radioligand [3H]5-amino-7-(2-phenylethyl)-2-(2-furyl)-pyrazolo[4,3-e]-1,2,4-triazolo [1,5-c] pyrimidine ([3H]SCH 58261). Saturation experiments revealed that [3H]SCH 58261 labels a single class of recognition sites with high affinity (Kd = 0.85 nM), limited capacity (apparent Bmax = 85 fmol/mg of protein) and good specific binding (about 60%). [3H]SCH 58261 binding was not modulated by either the divalent cation Mg(+2) or guanine nucleotides. In competition experiments, a series of both adenosine agonists and antagonists inhibited [3H]SCH 58261 binding to A2A platelet receptors with rank order of potency and affinity similar to those observed in rat striatal membranes with the same radioligand. This confirms that the platelet A2A receptor is similar to that labeled in the brain striatum. Binding data were also found to be in good agreement with the results from functional studies such as A2A agonist-induced stimulation of adenylate cyclase or platelet aggregation inhibition. The present findings indicate that [3H]SCH 58261 is the first radioligand available for the characterization of the A2A receptor subtype in platelets.
authors.affiliation: Schering-Plough Research Institute, Milan, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Dionisotti
authors.forename: S
authors.initials: S
authors.lastname: Ferrara
authors.forename: C
authors.initials: C
authors.lastname: Molta
authors.forename: C
authors.initials: C
authors.lastname: Zocchi
authors.forename: E
authors.initials: E
authors.lastname: Ongini
mesh_terms: D001667:Binding, Competitive; D001792:Blood Platelets; D002462:Cell Membrane / Q000378:metabolism; D002474:Cell-Free System; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D058906:Purinergic P1 Receptor Agonists; D058915:Purinergic P1 Receptor Antagonists; D011743:Pyrimidines / Q000737:chemistry / Q000378:metabolism*; D011869:Radioligand Assay; D018047:Receptors, Purinergic P1 / Q000378:metabolism*; D014230:Triazoles / Q000737:chemistry / Q000378:metabolism*
chemical_list: C098657:5-amino-7-(2-phenylethyl)-2-(2-furyl)pyrazolo(4,3-e)-1,2,4-triazolo(1,5-c)pyrimidine; D058906:Purinergic P1 Receptor Agonists; D058915:Purinergic P1 Receptor Antagonists; D011743:Pyrimidines; D018047:Receptors, Purinergic P1; D014230:Triazoles