Bioluminescence in G Protein-Coupled Receptors Drug Screening Using Nanoluciferase and Halo-Tag Technology

Methods Mol Biol. 2021:2268:137-147. doi: 10.1007/978-1-0716-1221-7_9.

Abstract

Here we describe the stepwise application of bioluminescence resonance energy transfer (BRET)-based conformational receptor biosensors to study GPCR activation in intact cells. This technology can be easily adopted to various plate reader devices and microtiter plate formats. Due to the high sensitivity of these BRET-based receptor biosensors and their ability to quantify simultaneously receptor activation/de-activation kinetics as well as compound efficacy and potency, these optical tools provide the most direct and unbiased approach to monitor GPCR activity in a high-throughput-compatible assay format, representing a novel promising tool for the discovery of potential GPCR therapeutics.

Keywords: Activation dynamics; Bioluminescence resonance energy transfer; Conformational biosensors; Drug screening; G protein-coupled receptors; Z-factor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bioluminescence Resonance Energy Transfer Techniques / methods*
  • Drug Evaluation, Preclinical / methods*
  • Fluorescent Dyes / chemistry*
  • HEK293 Cells
  • High-Throughput Screening Assays / methods*
  • Humans
  • Luciferases / metabolism*
  • Protein Conformation
  • Receptors, G-Protein-Coupled / chemistry
  • Receptors, G-Protein-Coupled / metabolism*

Substances

  • Fluorescent Dyes
  • Receptors, G-Protein-Coupled
  • Luciferases