Approaches to characterize and quantify oligomerization of GPCRs

Marsango, S., Varela, M. J. and Milligan, G. (2015) Approaches to characterize and quantify oligomerization of GPCRs. In: Filizola, M. (ed.) G Protein-Coupled Receptors in Drug Discovery. Series: Methods in Molecular Biology, 1335 (1335). Springer New York, pp. 95-105. ISBN 9781493929139 (doi: 10.1007/978-1-4939-2914-6_7)

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Abstract

Fluorescence resonance energy transfer (FRET) is an approach widely used to detect protein–protein interactions in live cells. This approach is based on the sensitization of an “acceptor” molecule by the energy transfer from a “donor” when there is an overlap between the emission spectrum of the “donor” and the excitation spectrum of the “acceptor” and close proximity between the two fluorophore species (in the region of 8 nm). Various methods exist to quantify FRET signals: here, we describe the application of homogeneous time-resolved FRET (htrFRET) combined with Tag-lite™ technology and its application to determine not only protein–protein interactions but also the capability of GPCR mutant variants to form homomers compared to the wild type GPCR within the plasma membrane of transfected cells.

Item Type:Book Sections
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Varela, Dr Maria Jose and Milligan, Professor Graeme and Marsango, Dr Sara
Authors: Marsango, S., Varela, M. J., and Milligan, G.
College/School:College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:Methods in Molecular Biology
Publisher:Springer New York
ISSN:1064-3745
ISSN (Online):1940-6029
ISBN:9781493929139
Copyright Holders:Copyright © 2016 Springer New York
First Published:First published in Methods in Molecular Biology 1335:95-105
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
656901The organisational structure of class A GPCRs: Implications for pharmacology, function and therapeutic regulationGraeme MilliganMedical Research Council (MRC)MR/L023806/1RI MOLECULAR CELL & SYSTEMS BIOLOGY
510631The organisational structure of class A GPCRs: implications for function and drug designGraeme MilliganMedical Research Council (MRC)G0900050RI NEUROSCIENCE & PSYCHOLOGY