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Guanine nucleotide binding sites, responsible for adenylate cyclase activation and carbamylcholine binding inhibition, show similar properties in rat heart membranes

par Waelbroeck, Magali ;Robberecht, Patrick ;Chatelain, Pierre ;Christophe, Jean
Référence Journal of cyclic nucleotide research, 7, 3, page (173-185)
Publication Publié, 1981

Article révisé par les pairs

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Résumé :

1/ In rat heart membranes, muscarinic receptors were shown to interact with guanine nucleotide binding sites closely related, if not identical, to those activating adenylate cyclase. The dose-effect curves of GTP, p[NH]ppG, and GTP gamma S for inhibition of carbamylcholine binding (measured by competition with [3H]QNB) and for adenylate cyclase activation (measured in the presence of isoproterenol) were parallel, at both 25 degrees C and 37 degrees C. 2/ Persistent activation of adenylate cyclase was obtained in heart membranes preincubated with p[NH]ppG or GTP gamma S then washed. The affinity for carbamylcholine was reduced after this pretreatment. The SO.5 of p[NH]ppG and GTP gamma S provoking persistent activation of adenylate cyclase and persistent inhibition of carbamylcholine binding were identical. Persistent inhibition of carbamylcholine binding was not additive with the inhibition observed when fresh nucleotide was added after washing. With p[NH]ppG, SO.5 values were unaffected by washing. With GTP gamma S, the SO.5 value for persistent activation of adenylate cyclase (i.e. after washing) and 330 times higher than that implementing activation (i.e. before washing). A similar change was observed when testing the SO.5 of GTP gamma S inhibition of carbamylcholine binding. This might reflect a partial release of GTP gamma S (but not of p[NH]ppG) from "spare" nucleotide binding sites during the washing period. 3/ Adenylate cyclase activity after maximal persistent activation was not increased when 0.1 mM guanine nucleotide, with or without 10 muM isoproterenol, was added to the incubation medium. In contrast, carbamylcholine binding was further decreased when fresh guanine nucleotide was added to the binding assay. This suggests that the proportion of "spare" nucleotide binding sites capable of activating the adenylate cyclase was higher than that capable to inhibit carbamylcholine binding, or that a second class of nucleotide binding sites (binding p[NH]ppG and GTP gamma S reversibly) was also able to inhibit carbamylcholine binding.