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Updated: Aug 16, 2026

Quantifying Agonist Activity at G Protein-coupled Receptors
Published on: December 26, 2011
Copper ions and diamide induce a high affinity guanine-nucleotide-insensitive state for muscarinic agonists
Abstract:
The binding capacity of [3H]-acetylcholine for muscarinic receptors of rat cerebral cortex membranes is increased in the presence of Cu2+ ions from 690 to 1320 fmol/mg protein with no significant change in affinity. Membranes treated with 50 microM Cu2+ and washed retain the increased binding capacity. Agonist binding in copper-treated membranes is insensitive to guanylylimidodiphosphate even at high concentrations (greater than 200 microM). Similar results were obtained when the sulfhydryl oxidizing agent, diamide (2 mM) was substituted for Cu2+ in the treatment of membranes. These data suggest the involvement of inter- or intra-molecular SH/S-S transitions in the interaction between the muscarinic receptor and a guanine nucleotide binding regulatory protein.
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