Some structural requirements which are important for the contractile activity of adenine nucleotides at P 2-purinergic receptors in the guinea-pig isolated vas deferens were examined. The consequences of deletions, substitutions and isosteric rearrangements in 1-N, C-2, C-8, N-9 and N 6 of adenine, in the ribose hydroxyls, and in the polyphosphate chain were examined. Several kinds of effects on activity were observed, including potentiation of responses, losses in activity, modifications of response duration, and transitions from biphasic concentration-response curves, which are characteristic of ATP, to monophasic curves. Generally, deletions and substitutions to adenine reduced activity. Ribose modifications were better-tolerated. The presence of 5′-phosphate or phosphorothioate moieties was required for maximum activity. A comparison of the present results with previous reports indicates that certain modifications to ATP result in similar effects in both the vas deferens and urinary bladder, which are opposite those which occur in the taenia coli.