PT - JOURNAL ARTICLE AU - Hartmut Berger AU - Girolamo Calo' AU - Erika Albrecht AU - Remo Guerrini AU - Michael Bienert TI - [Nphe<sup>1</sup>]NC(1-13)NH<sub>2</sub> Selectively Antagonizes Nociceptin/Orphanin FQ-Stimulated G-Protein Activation in Rat Brain DP - 2000 Aug 01 TA - Journal of Pharmacology and Experimental Therapeutics PG - 428--433 VI - 294 IP - 2 4099 - http://jpet.aspetjournals.org/content/294/2/428.short 4100 - http://jpet.aspetjournals.org/content/294/2/428.full SO - J Pharmacol Exp Ther2000 Aug 01; 294 AB - [Phe1psi(CH2-NH)Gly2]noc/OFQ(1-13)-amide {[F/G]NC(1-13)NH2} and acetyl-RYYRIK-amide (Ac-RYYRIK-NH2), two peptidic ligands of the nociceptin/orphanin FQ (noc/OFQ) receptor, have been shown to exert both agonist and antagonist activity in different in vitro and in vivo systems. This is despite the observation that both peptides competitively antagonized the coupling of the activated receptor to G-proteins in brain preparations, measured in GTPγ35S binding assays. In this study, [Nphe1]NC(1-13)-amide ([Nphe1]NC(1-13)NH2), a new noc/OFQ analog recently characterized as a pure and selective noc/OFQ receptor antagonist in several in vitro and in vivo assay systems, was shown to competitively inhibit the noc/OFQ-stimulated GTPγ35S binding to rat cerebral cortex membranes with pA2 of 7.76 (Schild analysis). This antagonism of noc/OFQ receptor G-protein coupling was selective because the peptide inhibited the noc/OFQ-evoked GTPγ35S binding to rat brain membranes but not that evoked by selective agonists of the μ-, δ-, and κ-opioid receptors. In rat cortical membranes, the effects of [F/G]NC(1-13)NH2 and Ac-RYYRIK-NH2 on the binding of GTPγ35S were clearly differentiated from the effect of [Nphe1]NC(1-13)NH2 when the concentration of GDP, competing with GTPγS for binding, was lowered from 100 μM (assay optimum) to 5 μM. At 5 μM GDP, the former peptides showed clear partial agonist activity, whereas [Nphe1]NC(1-13)NH2 did not. These data indicate that only [Nphe1]NC(1-13)NH2 was a pure antagonist of noc/OFQ receptor G-protein coupling. Furthermore, it is suggested that the variable behavior of [F/G]NC(1-13)NH2 and Ac-RYYRIK-NH2 (agonist, partial agonist, and antagonist) in different in vitro and in vivo systems may be explained by different partial GTP binding agonism and the existence of a GTP binding stimulus/response reserve (coupling reserve). The American Society for Pharmacology and Experimental Therapeutics