Quantitation of Plasma Membrane (G Protein-Coupled) Receptor Trafficking in Cultured Cells

Jim Wager-Miller; Ken Mackie

doi:10.1007/978-1-0716-2728-0_33

Quantitation of Plasma Membrane (G Protein-Coupled) Receptor Trafficking in Cultured Cells

Methods Mol Biol. 2023:2576:395-406. doi: 10.1007/978-1-0716-2728-0_33.

Authors

Jim Wager-Miller¹, Ken Mackie²

Affiliations

¹ Department of Psychological and Brain Sciences, Indiana University Bloomington, Bloomington, IN, USA.
² Department of Psychological and Brain Sciences, Indiana University Bloomington, Bloomington, IN, USA. kmackie@indiana.edu.

PMID: 36152205
DOI: 10.1007/978-1-0716-2728-0_33

Abstract

Measuring the functional behavior of G protein-coupled receptors (GPCRs) has been a major focus of academic and pharmaceutical research for many decades. These efforts have led to the development of many assays to measure the downstream effects of ligand binding on receptor activity. In this chapter, we describe an internalization/recycling assay that can be used to track changes in receptor number at the plasma membrane. Used in concert with other assays, this antibody-based technique can provide dynamic information on GPCR activation by receptor-specific ligands.

Keywords: Agonist; Antagonist; CB1 cannabinoid receptor (CB1); Epitope tag; G protein-coupled receptor (GPCR); G protein-coupled receptor kinase (GRK); Recycling.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Cell Membrane / metabolism
Cells, Cultured
GTP-Binding Proteins* / metabolism
Ligands
Receptors, G-Protein-Coupled* / metabolism

Substances

Ligands
Receptors, G-Protein-Coupled
GTP-Binding Proteins