Lipid-dependent GPCR dimerization

Alan D. Goddard, Patricia M. Dijkman, Roslin J. Adamson, Anthony Watts

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

Abstract

It has been widely demonstrated that G protein-coupled receptors (GPCRs) can form dimers both in vivo and in vitro, a process that has functional consequences. These receptor-receptor interactions take place within a phospholipid bilayer, yet, generally, little is known of the requirements for specific lipids that mediate the dimerization process. Studying this phenomenon in vivo is challenging due to difficulties in modulating the lipid content of cell membranes. Therefore, in this chapter, we describe techniques for reconstitution of GPCRs into model lipid bilayers of defined composition. The concentrations of specific lipids and sterols can be precisely controlled in these liposomes, as well as maintaining an appropriate lipid-protein ratio to avoid artifactual interactions. Receptor dimerization in this system is monitored via Förster resonance energy transfer (FRET), which requires the use of fluorescently labeled receptors. We therefore also include protocols for labeling with appropriate fluorophores and determining the apparent FRET efficiency, a measurement of the extent of receptor dimerization. Understanding the lipid dependence of GPCR dimerization will be key in understanding how this process is regulated in the dynamic heterogeneous environment of the cell membrane.

Original languageEnglish
Title of host publicationReceptor-receptor interactions
EditorsP. Michael Conn
Place of PublicationLondon (UK)
PublisherAcademic Press
Pages341-357
Number of pages17
Volume117
ISBN (Print)978-0-12-408143-7
DOIs
Publication statusPublished - 2013

Publication series

NameMethods in Cell Biology
PublisherAcademic Press
Volume117
ISSN (Print)0091-679X

Fingerprint

Dimerization
G-Protein-Coupled Receptors
Lipids
Energy Transfer
Lipid Bilayers
Sterols
Membrane Lipids
Liposomes
Phospholipids
Cell Membrane
Proteins

Keywords

  • dimer
  • FRET
  • G protein-coupled receptor (GPCR)
  • lipid dependence
  • liposome
  • NTS1

Cite this

Goddard, A. D., Dijkman, P. M., Adamson, R. J., & Watts, A. (2013). Lipid-dependent GPCR dimerization. In P. M. Conn (Ed.), Receptor-receptor interactions (Vol. 117, pp. 341-357). (Methods in Cell Biology; Vol. 117). London (UK): Academic Press. https://doi.org/10.1016/B978-0-12-408143-7.00018-9
Goddard, Alan D. ; Dijkman, Patricia M. ; Adamson, Roslin J. ; Watts, Anthony. / Lipid-dependent GPCR dimerization. Receptor-receptor interactions. editor / P. Michael Conn. Vol. 117 London (UK) : Academic Press, 2013. pp. 341-357 (Methods in Cell Biology).
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Goddard, AD, Dijkman, PM, Adamson, RJ & Watts, A 2013, Lipid-dependent GPCR dimerization. in PM Conn (ed.), Receptor-receptor interactions. vol. 117, Methods in Cell Biology, vol. 117, Academic Press, London (UK), pp. 341-357. https://doi.org/10.1016/B978-0-12-408143-7.00018-9

Lipid-dependent GPCR dimerization. / Goddard, Alan D.; Dijkman, Patricia M.; Adamson, Roslin J.; Watts, Anthony.

Receptor-receptor interactions. ed. / P. Michael Conn. Vol. 117 London (UK) : Academic Press, 2013. p. 341-357 (Methods in Cell Biology; Vol. 117).

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

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Goddard AD, Dijkman PM, Adamson RJ, Watts A. Lipid-dependent GPCR dimerization. In Conn PM, editor, Receptor-receptor interactions. Vol. 117. London (UK): Academic Press. 2013. p. 341-357. (Methods in Cell Biology). https://doi.org/10.1016/B978-0-12-408143-7.00018-9