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
; 130(5): 1156-64, 2000 Jul.
Article
in En
| MEDLINE
| ID: mdl-10882402
ABSTRACT
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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Receptors, Purinergic P1
/
Cyclic AMP
/
GTP-Binding Proteins
Language:
En
Journal:
Br J Pharmacol
Year:
2000
Document type:
Article
Affiliation country:
United States