Regulation of muscarinic agonist binding by cations and guanine nucleotides

EDTA treatment of membrane preparations from rat brain and myocardium reduced the relative proportion of superhigh and high affinity binding sites for muscarinic agonists by up to 60%. This effect was partially or completely reversed by millimolar concentrations of Mg2+ or Mn2+. A number of multivalent metal cations gave qualitatively similar effects, yielding stimulation of agonist binding at low concentrations but inhibition at higher concentrations. The divalent cation-linked subpopulation of muscarinic agonist binding sites identified in this study appears to be the primary target for guanine nucleotide inhibition.