title: Unlocking the secrets of the gatekeeper: methods for stabilizing and crystallizing GPCRs.
abstract: G-protein coupled receptors (GPCRs) are integral membrane cell surface receptors with key roles in mediating the cellular responses to a wide range of biologically relevant molecules including hormones, neurotransmitters and importantly the majority of currently available drugs. The first high-resolution, X-ray crystallographic structure of a GPCR, that of rhodopsin, was obtained in 2000. It took a further seven years for the next structure, that of the β2 adrenergic receptor. Remarkably, at the time of writing, there have been an astonishing 18 further independent high-resolution GPCR structures published in the last five years (overall total of 68 structures in different conformations or bound to different ligands). Of particular note is the recent structure of the β2 adrenergic receptor in complex with its cognate heterotrimeric G-protein revealing for the first time molecular details of the interaction between a GPCR and the complete G-protein. Together these structures have provided unprecedented detail into the mechanism of action of these incredibly important proteins. This review describes several key methodological advances that have made such extraordinarily fast progress possible.
authors.affiliation: Department of Life Sciences, Imperial College London, SW7 2AZ, UK.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bertheleme
authors.forename: Pil Seok
authors.initials: PS
authors.lastname: Chae
authors.forename: Shweta
authors.initials: S
authors.lastname: Singh
authors.forename: Danuta
authors.initials: D
authors.lastname: Mossakowska
authors.forename: Michael M
authors.initials: MM
authors.lastname: Hann
authors.forename: Kathrine J
authors.initials: KJ
authors.lastname: Smith
authors.forename: Julia A
authors.initials: JA
authors.lastname: Hubbard
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Dowell
authors.forename: Bernadette
authors.initials: B
authors.lastname: Byrne
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D007128:Immunoglobulin Fragments / Q000378:metabolism; D008958:Models, Molecular; D016296:Mutagenesis; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D007128:Immunoglobulin Fragments; D043562:Receptors, G-Protein-Coupled
keywords: Antibody; Detergent; G-protein coupled receptor; Genetic modification; Stabilization; X-ray crystallography