RECIPROCAL MODULATION OF AGONIST AND ANTAGONIST BINDING TO A1 ADENOSINE RECEPTORS BY GUANINE-NUCLEOTIDES IS MEDIATED VIA A PERTUSSIS TOXIN-SENSITIVE G-PROTEIN

Google Scholar
  • 1 September 1988
    • journal article
    • research article
    • Vol. 246  (3) , 1194-1200
Abstract
In the present study, we have characterized the effects of guanine nucleotides on agonist and antagonist binding to A1 adenosine receptors, which mediate inhibition of adenylate cyclase via the inhibitory G protein (Gi) in adipocytes. Our data indicate that guanosine-triphosphate (GTP) and guanyl-5''-yl imididodiphospahte (Gpp(NH)p) enhance the binding of 8-[ GDP > GDP.beta.S = cGMP, which paralleled their ability to inhibit forskolin-stimulated adenylate cyclase activity. Saturation isotherms performed in the absence and presence of GTP and Gpp(NH)p indicate that the guanine nucleotide decreased the equilibrium dissociation constant (KD) but had no effect on the maximal binding (Bmax) of [3H]XAC. In contrast, Gpp(NH)p decreased agonist binding as manifested by a decrease in the percentage of A1 adenosine receptors in the agonist high affinity state (from 81% of 27%) without changing the high (KH) and low (KL) affinity constants. Kinetic experiments conducted to assess the effect of guanine nucleotide on [3H]XAC binding parameters demonstrate that Gpp(NH)p enhanced the observed rate of association (Kobs) of the radioligand with the receptor by 2-fold but had no effect on the rate of dissociation (K-1) of the radioligand-receptor complex. Taken together, these results that guanine nucleotides reciprocally affect agonist and antagonist binding to the A1 adenosine receptor. Enhancement in antagonist binding affinity is dependent on a pertussis toxin-sensitive G protein, presumably G1, which apparently can, under the influence of guanine nucleotides, induce a conformation of the receptor possessing higher affinity for the antagonist.

This publication has 19 references indexed in Scilit: