NanoBRET Approaches to Study Ligand Binding to GPCRs and RTKs

Trends Pharmacol Sci. 2018 Feb;39(2):136-147. doi: 10.1016/j.tips.2017.10.006. Epub 2017 Nov 10.

Abstract

Recent advances in the development of fluorescent ligands for G-protein-coupled receptors (GPCRs) and receptor tyrosine kinase receptors (RTKs) have facilitated the study of these receptors in living cells. A limitation of these ligands is potential uptake into cells and increased nonspecific binding. However, this can largely be overcome by using proximity approaches, such as bioluminescence resonance energy transfer (BRET), which localise the signal (within 10nm) to the specific receptor target. The recent engineering of NanoLuc has resulted in a luciferase variant that is smaller and significantly brighter (up to tenfold) than existing variants. Here, we review the use of BRET from N-terminal NanoLuc-tagged GPCRs or a RTK to a receptor-bound fluorescent ligand to provide quantitative pharmacology of ligand-receptor interactions in living cells in real time.

Keywords: G-protein-coupled receptor; NanoBRET; NanoLuc luciferase; bioluminescence; ligand binding; receptor tyrosine kinase.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Bioluminescence Resonance Energy Transfer Techniques / methods*
  • Humans
  • Ligands
  • Protein Binding
  • Receptor Protein-Tyrosine Kinases / chemistry*
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Receptors, G-Protein-Coupled / chemistry*
  • Receptors, G-Protein-Coupled / metabolism
  • Single Molecule Imaging / methods*

Substances

  • Ligands
  • Receptors, G-Protein-Coupled
  • Receptor Protein-Tyrosine Kinases