Biophysical characterization of G-protein coupled receptor–peptide ligand bindingThis paper is one of a selection of papers published in a Special Issue entitled CSBMCB 53rd Annual Meeting — Membrane Proteins in Health and Disease, and has undergone the Journal’s usual peer review process.

Abstract

G-protein coupled receptors (GPCRs) are ubiquitous membrane proteins allowing intracellular response to extracellular factors that range from photons of light to small molecules to proteins. Despite extensive exploitation of GRCRs as therapeutic targets, biophysical characterization of GPCR-ligand interactions remains challenging. In this minireview, we focus on techniques which have been successfully employed for structural and biophysical characterization of peptide ligands binding to their cognate GPCRs. The techniques reviewed include solution-state nuclear magnetic resonance (NMR) spectroscopy; solid-state NMR; X-ray diffraction; fluorescence spectroscopy and single molecule fluorescence methods; flow cytometry; surface plasmon resonance; isothermal titration calorimetry; and, atomic force microscopy. The goal herein is to provide a cohesive starting point to allow selection of techniques appropriate to the elucidation of a given GPCR-peptide interaction.

Arginine-, d-arginine-vasopressin, and their inverso analogues in micellar and liposomic models of cell membrane: CD, NMR, and molecular dynamics studies

“…There is evidence to supporting the membrane-bound pathway for the interaction of a ligand with its cognate GPCRs (Moroder et al 1993 ; Langelaan and Rainey 2010 ; Langelaan et al 2011 ). In this mechanism, adsorption of the ligand to the cell membrane is followed by a two-dimensional diffusion process, whereby the ligand binds to and activates the receptor (Schwyzer 1995 ; Mierke and Giragossian 2001 ).…”

Mass Spectrometry-based Proteomics of Human Cannabinoid Receptor 2: Covalent Cysteine 6.47(257)-Ligand Interaction Affording Megagonist Receptor Activation

“…46 These aggregate data invite speculation that covalent binding of AM-841 to hCB2R C6.47(257) may induce and/or stabilize conformational change in this receptor region to elicit structural features highly advantageous to hCB2R activation. Successful crystallization of AM-841-liganded hCB2R followed by X-ray analysis of the complex would be an interesting, if at present technologically elusive, 3,47 means of characterizing what such a highly-activated hCB2R structure might represent.…”

Structure‐function studies with G protein‐coupled receptors as a paradigm for improving drug discovery and development of therapeutics

“…upon addition of ligands. Many groups have used this technique to calculate the dissociation constant or to ensure that the purified receptor retains ligand-binding activity [96]. Environmentally sensitive probes and intrinsic fluorescence have also been used to monitor the interaction between receptors and G proteins; fluorescence was measured and conformational changes and binding rates were observed with various ligands and detergent and lipid compositions [97-99].…”