Differential A(1)-adenosine receptor reserve for inhibition of cyclic AMP accumulation and G-protein activation in DDT(1) MF-2 cells

Br J Pharmacol. 2000 Jul;130(5):1156-64. doi: 10.1038/sj.bjp.0703405.


1. The A(1)-adenosine receptor (A(1)AdoR) reserve for N(6)-cyclopentyladenosine (CPA) mediated inhibition of (-)isoprenaline stimulated cyclic AMP accumulation and stimulation of [(35)S]-guanosine-5'-O-(thiotriphosphate) (GTPgammaS) binding, a measure of guanine nucleotide binding protein (G-protein) activation, was determined in DDT(1) MF-2 cells. 2. Inactivation of the A(1)AdoRs with the chemoreactive ligand 8-cyclopentyl-3-[3-[[4-(fluorosulphonyl)benzoyl]oxy]propyl]-1-p ropylx anthine (FSCPX) caused a progressive rightward shift of the concentration-response curves for CPA to inhibit cyclic AMP accumulation, with a maximum of 10 fold increase in the EC(50) value. In contrast, inactivation of A(1)AdoR's caused only a 1.7 fold rightward shift in the CPA concentration-response for stimulation of [(35)S]-GTPgammaS binding. 3. The A(1)AdoR occupancy-response relationship for CPA inhibition of cyclic AMP accumulation was hyperbolic with 43% receptor occupancy required to elicit the maximal response, i.e. a 57% A(1)AdoR reserve. In contrast, the A(1)AdoR occupancy-response relationship for CPA mediated stimulation of [(35)S]-GTPgammaS binding was linear indicating little or no receptor reserve for G-protein activation. The relationship between CPA stimulation of [(35)S]-GTPgammaS binding and cyclic AMP inhibition was also hyperbolic with 44% G-protein activation sufficient to cause maximal inhibition. 4. The data suggest that the A(1)AdoR reserve for CPA mediated inhibition of cyclic AMP accumulation occurs at the level of G-protein interaction with adenylyl cyclase. However, each A(1)AdoR appears to activate a constant fraction of the total G-protein population suggesting signal amplification at the receptor-G-protein level which may also contribute to the receptor reserve for CPA.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine / analogs & derivatives
  • Adenosine / pharmacology
  • Cells, Cultured
  • Cyclic AMP / antagonists & inhibitors*
  • Dose-Response Relationship, Drug
  • GTP-Binding Proteins / physiology*
  • Guanosine 5'-O-(3-Thiotriphosphate) / metabolism
  • Muscle, Smooth / metabolism
  • Receptors, Purinergic P1 / physiology*
  • Xanthines / pharmacology


  • Receptors, Purinergic P1
  • Xanthines
  • 8-cyclopentyl-3-(3-((4-(fluorosulfonyl)benzoyl)oxy)propyl)-1-propylxanthine
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • N(6)-cyclopentyladenosine
  • Cyclic AMP
  • GTP-Binding Proteins
  • Adenosine