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Updated: Jul 28, 2026

Quantifying Agonist Activity at G Protein-coupled Receptors
Published on: December 26, 2011
Assays for enhanced activity of low efficacy partial agonists at the D(2) dopamine receptor
H Lin1, S G N Saisch, P G Strange
1School of Pharmacy, University of Reading, Whiteknights, Reading, Berkshire, UK.
Background And Purpose:
Low efficacy partial agonists at the D2 dopamine receptor may be useful for treating schizophrenia. In this report we describe a method for assessing the efficacy of these compounds based on stimulation of [35S]GTPgammaS binding.
Experimental Approach:
Agonist efficacy was assessed from [(35)S]GTPgammaS binding to membranes of CHO cells expressing D2 dopamine receptors in buffers with and without Na+. Effects of Na+ on receptor/G protein coupling were assessed using agonist/[3H]spiperone competition binding assays.
Key Results:
When [35S]GTPgammaS binding assays were performed in buffers containing Na+, some agonists (aripiprazole, AJ-76, UH-232) exhibited very low efficacy whereas other agonists exhibited measurable efficacy. When Na+ was substituted by N-methyl D-glucamine, the efficacy of all agonists increased (relative to that of dopamine) but particularly for aripiprazole, aplindore, AJ-76, (-)-3-PPP and UH-232. In ligand binding assays, substitution of Na+ by N-methyl D-glucamine increased receptor/G protein coupling for some agonists -. aplindore, dopamine and (-)-3-PPP - but for aripiprazole, AJ-76 and UH-232 there was little effect on receptor/G protein coupling.
Conclusions And Implications:
Substitution of Na+ by NMDG increases sensitivity in [(35)S]GTPgammaS binding assays so that very low efficacy agonists were detected clearly. For some agonists the effect seems to be mediated via enhanced receptor/G protein coupling whereas for others the effect is mediated at another point in the G protein activation cycle. AJ-76, aripiprazole and UH-232 seem particularly sensitive to this change in assay conditions. This work provides a new method to discover these very low efficacy agonists.
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