The structure-activity relationships for a variety of adenine nucleotide analogues at P 2x -and P 2Y -purinoceptors were investigated. Compounds formed by structural modifications of the ATP molecule including substitutions of the purine ring (C2, C8, N1, and N 6 -substituents, and a uridine base instead of adenine), the ribose moiety (2′ and 3′-positions), and the triphosphate group (lower phosphates, bridging oxygen substitution, and cyclization) were prepared. Pharmacological activity at P 2Y -purinoceptors was assayed in the guinea pig taenia coli, endothelial cells of the rabbit aorta, smooth muscle of the rabbit mesenteric artery, and turkey erythrocyte membranes. Activity at P 2X -purinoceptors was assayed in the rabbit saphenous artery and the guinea-pig vas deferens and urinary bladder. Some of the analogues displayed selectivity, or even specificity, for either the P 2X -or the P 2Y -purinoceptors. Certain analogues displayed selectivity or specificity within the P 2X -or P 2Y -purinoceptor superfamilies, giving hints about possible subclasses. For example, 8-(6-aminohexylamino)ATP and 2′,3′-isopropylidene-AMP were selective for endothelial Pzypurinoceptors over P 2Y -purinoceptors in the guinea pig taenia coli, rabbit aorta, and turkey erythrocytes. These compounds were both inactive at P 2X -purinoceptors. The potent agonist N 6 -methyl ATP and the somewhat less potent agonist 2′-deoxy-ATP were selective for P 2Y -purinoceptors in the guinea pig taenia coli, but were inactive at P 2X -purinoceptors and the vascular P 2Y -purinoceptors. 3′-Benzylamino-3′-deoxyATP was very potent at the P 2X -purinoceptors in the guinea pig vas deferens and bladder, but not in the rabbit saphenous artery and was inactive at P 2Y receptors. These data suggest that specific compounds can be developed that can be utilized to activate putative subtypes of the P 2X -and P 2Y -purinoceptor classes.