Detection of genome-edited and endogenously expressed G protein-coupled receptors.

Mark Soave,Carl W White,Stephen J Hill,Leigh A Stoddart,Joëlle Goulding,Laura E Kilpatrick,Stephen J Briddon

doi:10.1111/febs.15729

Abstract

G protein‐coupled receptors (GPCRs) are the largest family of membrane receptors and major targets for FDA‐approved drugs. The ability to quantify GPCR expression and ligand binding characteristics in different cell types and tissues is therefore important for drug discovery. The advent of genome editing along with developments in fluorescent ligand design offers exciting new possibilities to probe GPCRs in their native environment. This review provides an overview of the recent technical advances employed to study the localisation and ligand binding characteristics of genome‐edited and endogenously expressed GPCRs.

Highlights

Gprotein-coupled receptors (GPCRs) are membrane proteins characterised by seven transmembrane-spanning domains, an extracellular N terminus and an intracellular C terminus
Improvements in fluorescent ligand design will result in more studies describing the subcellular GPCR localisation and how this may change in response to cellular stress
These approaches could be used in conjunction with advanced microscopy (Table 2), such as single-molecule tracking or fluorescence correlation spectroscopy (FCS), to probe the role of GPCR dimerisation and organisation into higher order oligomers in their native environment

Summary

Introduction

Gprotein-coupled receptors (GPCRs) are membrane proteins characterised by seven transmembrane-spanning domains, an extracellular N terminus and an intracellular C terminus. Ligand-directed labelling has been used to label endogenously expressed adenosine A2A receptors and l opioid receptors. The adenosine A2A receptor is a target for cancer immunotherapy as it is highly expressed on immune cells [111]. Expressed A2AR was detected using FACS analysis on human monocyte-derived macrophages and visualised on a human breast cancer cell line using confocal microscopy [109].

Results

Conclusion