[³⁵S]GTPγS binding in G protein-coupled receptor assays
Date
2009
Authors
Cooper, T.
McMurchie, E.
Leifert, W.
Editors
Lundstrom, K.
Chiu, M.
Chiu, M.
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Journal article
Citation
Methods in Molecular Biology, 2009; 552:143-151
Statement of Responsibility
Tamara Cooper, Edward J. McMurchie and Wayne R. Leifert
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Abstract
The [³⁵S]GTPγS binding assay to measure G protein activation following agonist binding to G protein-coupled receptors (GPCRs) remains a powerful molecular technique to substantiate traditional pharmacological values of potency, efficacy, and affinity. The method described uses membrane preparations of the α2A-adrenergic receptor and purified G protein subunits expressed in Sf9 cells, reconstituted into a functional signaling system. This technology is generic and could be used with other GPCRs to demonstrate initial signaling events following receptor activation. Agonist-stimulated [³⁵S]GTPγS binding is measured in a 96-well plate format using scintillation counting.
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Copyright 2009 Springer