Surveying GPCR solubilisation conditions using surface plasmon resonance

Iva Hopkins Navratilova (Lead / Corresponding author), Tonia Aristotelous, Louise E Bird, Andrew L Hopkins

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Biophysical screening techniques, such as surface plasmon resonance, enable detailed kinetic analysis of ligands binding to solubilised G-protein coupled receptors. The activity of a receptor solubilised out of the membrane is crucially dependent on the environment in which it is suspended. Finding the right conditions is challenging due to the number of variables to investigate in order to determine the optimum solubilisation buffer for any given receptor. In this study we used surface plasmon resonance technology to screen a variety of solubilisation conditions including buffers and detergents for two model receptors: CXCR4 and CCR5. We tested 950 different combinations of solubilisation conditions for both receptors. The activity of both receptors was monitored by using conformation dependent monoclonal antibodies and the binding of small molecule ligands. Despite both receptors belonging to the chemokine receptor family they show some differences in their preference for solubilisation conditions that provide the highest level of binding for both the conformation dependent antibodies and small molecules. The study described here is focused not only on finding the best solubilisation conditions for each receptor, but also on factors that determine the sensitivity of the assay for each receptor. We also suggest how these data about different buffers and detergents can be used as a guide for selecting solubilisation conditions for other membrane proteins.

Original languageEnglish
Pages (from-to)23-34
Number of pages12
JournalAnalytical Biochemistry
Volume556
Early online date15 Jun 2018
DOIs
Publication statusPublished - 1 Sep 2018

Fingerprint

Surface Plasmon Resonance
Surveying
Surface plasmon resonance
Buffers
Detergents
Conformations
CCR5 Receptors
CXCR4 Receptors
Ligands
Molecules
Chemokine Receptors
G-Protein-Coupled Receptors
Assays
Screening
Membrane Proteins
Monoclonal Antibodies
Technology
Membranes
Kinetics
Antibodies

Keywords

  • Detergents
  • GPCRs
  • Screening
  • Solubilisation conditions
  • Surface plasmon resonance

Cite this

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abstract = "Biophysical screening techniques, such as surface plasmon resonance, enable detailed kinetic analysis of ligands binding to solubilised G-protein coupled receptors. The activity of a receptor solubilised out of the membrane is crucially dependent on the environment in which it is suspended. Finding the right conditions is challenging due to the number of variables to investigate in order to determine the optimum solubilisation buffer for any given receptor. In this study we used surface plasmon resonance technology to screen a variety of solubilisation conditions including buffers and detergents for two model receptors: CXCR4 and CCR5. We tested 950 different combinations of solubilisation conditions for both receptors. The activity of both receptors was monitored by using conformation dependent monoclonal antibodies and the binding of small molecule ligands. Despite both receptors belonging to the chemokine receptor family they show some differences in their preference for solubilisation conditions that provide the highest level of binding for both the conformation dependent antibodies and small molecules. The study described here is focused not only on finding the best solubilisation conditions for each receptor, but also on factors that determine the sensitivity of the assay for each receptor. We also suggest how these data about different buffers and detergents can be used as a guide for selecting solubilisation conditions for other membrane proteins.",
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Surveying GPCR solubilisation conditions using surface plasmon resonance. / Navratilova, Iva Hopkins (Lead / Corresponding author); Aristotelous, Tonia; Bird, Louise E; Hopkins, Andrew L.

In: Analytical Biochemistry, Vol. 556, 01.09.2018, p. 23-34.

Research output: Contribution to journalArticle

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