tailieunhanh - Báo cáo khoa học: Methods to monitor the quaternary structure of G protein-coupled receptors

A wide range of approaches has been applied to examine the quaternary structure of G protein-coupled receptors, the basis of such protein–protein interactions and how such interactions might modulate the pharmacology and function of these receptors. These include coimmunoprecipitation, var-ious adaptations of resonance energy transfer techniques, functional com-plementation studies and the analysis of ligand-binding data. | iFEBS Journal MINIREVIEW Methods to monitor the quaternary structure of G protein-coupled receptors Graeme Milligan1 and Michel Bouvier2 1 Molecular Pharmacology Group Division of Biochemistry and Molecular Biology Institute of Biomedicaland Life Sciences University of Glasgow UK 2 Biochemistry Universite de Montreal Quebec Canada Keywords dimerization functional reconstitution G protein coupled receptor immunoprecipitation resonance energy transfer Correspondence G. Milligan Davidson Building University of Glasgow Glasgow G12 8QQ Scotland UK Fax 44 141330 4620 Tel 44 141330 5557 E-mail Received 16 February 2005 accepted 4 April 2005 A wide range of approaches has been applied to examine the quaternary structure of G protein-coupled receptors the basis of such protein-protein interactions and how such interactions might modulate the pharmacology and function of these receptors. These include coimmunoprecipitation various adaptations of resonance energy transfer techniques functional complementation studies and the analysis of ligand-binding data. Each of the available techniques has limitations that restrict interpretation of the data. However taken together they provide a coherent body of evidence indicating that many if not all G protein-coupled receptors exist and function as dimer oligomers. Herein we assess the widely applied techniques and discuss the relative benefits and limitations of these approaches. doi Introduction In recent times the view that G protein-coupled receptors GPCRs are single polypeptides that function as isolated monomers has been challenged and largely supplanted by results consistent with the existence of GPCRs as dimers or higher-order oligomers. Data in support of GPCRs being able to form dimers or oligomers had been scattered throughout the literature 1 . However at least in part because expression of a single GPCR cDNA in heterologous cell systems generally resulted in .