Ehlert_1980_J.Supramol.Struct_14_149

The regulation of muscarinic receptor binding by guanine nucleotides and N-ethylmaleimide (NEM) was investigated using the agonist ligand, [3H] cis methyldioxolane ([3H]CD). Characterization studies on rat forebrain homogenates show that [3H]CD binding was linear with tissue concentration and was unaffected by a change in pH from 5.5 to 8.0. The regional variation in [3H]CD binding in the rat brain correlated generally wit [3H](-)3-quinuclidinyl benzilate ([3H](-)QNB) binding, although the absolute variation in binding was somewhat less. At a concentration of 100 microM, the GTP analogue, guanyl-5'-yl imidodiphosphate [Gpp(NH)p], caused a 43-77% inhibition of [3H]CD binding in the corpus striatum, ileum, and heart. The results of binding studies using several Gpp(NH)p concentrations demonstrated that the potency of this guanine nucleotide for inhibition of [3H]CD binding, Gpp(NH)p caused an increase in (3H](-)QNB binding in the heart heart and ileum and no change in [3H](-)QNB binding in the corpus striatum. When measured by competitive inhibition of [3H](-)QNB binding to the longitudinal muscle of the ileum, Gpp(NH)p (100 microM) caused an increase in the IC50 values of a series of agonists in a manner that was correlated with the efficacy of these compounds. The results of binding studies on NEM treated forebrain homogenates revealed an enhancement of [3H]CD binding by NEM.