Analysis of function of receptor-G-protein and receptor-RGS fusion proteins

Ward, R.J. and Milligan, G. (2004) Analysis of function of receptor-G-protein and receptor-RGS fusion proteins. Methods in Molecular Biology, 259, pp. 225-247. (PMID:15250496)

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Abstract

Fusion constructs between G-protein-coupled receptors and G-protein alpha-subunits have been used to examine a variety of aspects of the functioning of these signaling systems. Here we describe some of the various techniques and methods used in detail. The process of fusing the two components, the receptor and the G-protein alpha-subunit, result in a construct that acts as an agonist stimulated GTPase enzyme whose expression level can be accurately determined by radioligand binding. The effects of different ligands, mutations, and other signaling system components can thus be analyzed by a series of relatively simple assays. Recently we have begun to supplement this approach with the use of fusions between G-protein-coupled receptors and regulator of G-protein signaling proteins to examine the function of this important group of signaling molecules.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Ward, Dr Richard and Milligan, Professor Graeme
Authors: Ward, R.J., and Milligan, G.
College/School:College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
Journal Name:Methods in Molecular Biology
Publisher:Humana Press Inc
ISSN:1064-3745
ISSN (Online):1940-6029

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
325011Post transitional modification and the function of regulator of G protein signallingGraeme MilliganBiotechnology and Biological Sciences Research Council (BBSRC)17/C17789Institute of Neuroscience and Psychology