| Title: | Fluorescent approaches for understanding interactions of ligands with G protein coupled receptors |
| Author(s): | Sridharan R; Zuber J; Connelly SM; Mathew E; Dumont ME; |
| Address: | "Department of Biochemistry and Biophysics, P.O. Box 712, University of Rochester Medical Center, Rochester, NY 14642, USA" |
| DOI: | 10.1016/j.bbamem.2013.09.005 |
| ISSN/ISBN: | 0006-3002 (Print) 0006-3002 (Linking) |
| Abstract: | "G protein coupled receptors are responsible for a wide variety of signaling responses in diverse cell types. Despite major advances in the determination of structures of this class of receptors, the underlying mechanisms by which binding of different types of ligands specifically elicits particular signaling responses remain unclear. The use of fluorescence spectroscopy can provide important information about the process of ligand binding and ligand dependent conformational changes in receptors, especially kinetic aspects of these processes that can be difficult to extract from X-ray structures. We present an overview of the extensive array of fluorescent ligands that have been used in studies of G protein coupled receptors and describe spectroscopic approaches for assaying binding and probing the environment of receptor-bound ligands with particular attention to examples involving yeast pheromone receptors. In addition, we discuss the use of fluorescence spectroscopy for detecting and characterizing conformational changes in receptors induced by the binding of ligands. Such studies have provided strong evidence for diversity of receptor conformations elicited by different ligands, consistent with the idea that GPCRs are not simple on and off switches. This diversity of states constitutes an underlying mechanistic basis for biased agonism, the observation that different stimuli can produce different responses from a single receptor. It is likely that continued technical advances will allow fluorescence spectroscopy to play an important role in continued probing of structural transitions in G protein coupled receptors. This article is part of a Special Issue entitled: Structural and biophysical characterisation of membrane protein-ligand binding" |
| Keywords: | "Amino Acid Sequence Fluorescent Dyes/chemistry Kinetics Ligands Molecular Sequence Data Protein Binding Receptors, G-Protein-Coupled/chemistry/*metabolism X-Ray Diffraction Fret Fluorescence Fluorescence Resonance Energy Transfer G protein coupled recepto;" |
| Notes: | "MedlineSridharan, Rajashri Zuber, Jeffrey Connelly, Sara M Mathew, Elizabeth Dumont, Mark E eng R01 GM059357/GM/NIGMS NIH HHS/ R01 GM084083/GM/NIGMS NIH HHS/ U54 GM074899/GM/NIGMS NIH HHS/ U54 GM094611/GM/NIGMS NIH HHS/ U54 GM09461/GM/NIGMS NIH HHS/ Research Support, N.I.H., Extramural Review Netherlands 2013/09/24 Biochim Biophys Acta. 2014 Jan; 1838(1 Pt A):15-33. doi: 10.1016/j.bbamem.2013.09.005. Epub 2013 Sep 18" |
