Assays 1. GTPγS binding assays are performed according to the method of Sternweis and Robishaw.13 Samples are diluted with 10 mM Na–HEPES, pH 8.0, 1 mM EDTA, 1 mM DTT, 0.1% Thesit, so that the concentration is about 0.5 mg/ml. One to 5 μg of protein is included in each sam...
(2009). [35S]GTPgammaS binding in G protein-coupled receptor assays. Methods in molecular biology 552, 143-151.Cooper, T., McMurchie, E.J., and Leifert, W.R. (2009) [35S]GTPgammaS binding in G protein- coupled receptor assays. Methods Mol. Biol. 552, 143-151...
In this review I consider assays for G protein-coupled receptor (GPCR) activity based on the binding of labelled analogues of GTPgammaS ([(35) S]GTPgammaS or Eu-GTPgammaS) to G proteins in tissues (GTPgammaS binding assays). Such assays provide convenient measures of GPCR activity close...
Such assays provide convenient measures of GPCR activity close to the receptor in the signalling cascade. In order to set up a GTPgammaS binding assay, the requirements of the assay must be considered. These are tissue source, GTPgammaS analogue, G protein, GDP,...
Guanosine 5'-O-(3-[[sup.35]S]thio)triphosphate ([[sup.35]S]GTP[gamma]S) binding assays were established and utilized as a reliable and high-capacity functional assay for determining antagonist and inverse agonist pharmacological parameters of novel ...
The level of intrinsic activity for noribogaine in these assays was comparable to the full agonists DAMGO and morphine. In contrast, ibogaine had no significant effect on [35S]GTPgammaS binding over a similar concentration range. The efficacy of noribogaine as a full mu-opioid agoni...
Agonist responses were concentration-related, and highest efficacies were obtained in the hippocampus, thus paralleling both the brain regional distribution and agonist efficacies previously observed in phosphoinositide hydrolysis assays. These findings suggest that D1-like receptor conformations that mediate ...
Agonist-stimulated G[alpha]-GTP binding reflects a very early event in the activation of a GPCR signaling cascade, offering a significant advantage over the more distal events monitored by most reporter gene assays. Upon interaction of an agonist and a G[alph...