title: Methods used to study the oligomeric structure of G-protein-coupled receptors.
abstract: G-protein-coupled receptors (GPCRs), which constitute the largest family of cell surface receptors, were originally thought to function as monomers, but are now recognized as being able to act in a wide range of oligomeric states and indeed, it is known that the oligomerization state of a GPCR can modulate its pharmacology and function. A number of experimental techniques have been devised to study GPCR oligomerization including those based upon traditional biochemistry such as blue-native PAGE (BN-PAGE), co-immunoprecipitation (Co-IP) and protein-fragment complementation assays (PCAs), those based upon resonance energy transfer, FRET, time-resolved FRET (TR-FRET), FRET spectrometry and bioluminescence resonance energy transfer (BRET). Those based upon microscopy such as FRAP, total internal reflection fluorescence microscopy (TIRFM), spatial intensity distribution analysis (SpIDA) and various single molecule imaging techniques. Finally with the solution of a growing number of crystal structures, X-ray crystallography must be acknowledged as an important source of discovery in this field. A different, but in many ways complementary approach to the use of more traditional experimental techniques, are those involving computational methods that possess obvious merit in the study of the dynamics of oligomer formation and function. Here, we summarize the latest developments that have been made in the methods used to study GPCR oligomerization and give an overview of their application.
authors.affiliation: Faculty of Environmental Science and Engineering, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China.|First People's Hospital of Yunnan Province, Kunming, Yunnan, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Guo
authors.affiliation: Faculty of Environmental Science and Engineering, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China.
authors.forename: Su
authors.initials: S
authors.lastname: An
authors.affiliation: Molecular Pharmacology Group, Institute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, Scotland, U.K.
authors.forename: Richard
authors.initials: R
authors.lastname: Ward
authors.affiliation: Faculty of Environmental Science and Engineering, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Faculty of Environmental Science and Engineering, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Faculty of Environmental Science and Engineering, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China.
authors.forename: Xiao-Xi
authors.initials: XX
authors.lastname: Guo
authors.affiliation: Faculty of Environmental Science and Engineering, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Hao
authors.affiliation: Faculty of Environmental Science and Engineering, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China tianruixu@kmust.edu.cn.
authors.forename: Tian-Rui
authors.initials: TR
authors.lastname: Xu
mesh_terms: D000818:Animals; D018360:Crystallography, X-Ray / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D055503:Protein Multimerization / Q000502:physiology*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*
chemical_list: D043562:Receptors, G-Protein-Coupled
keywords: FRET technology; GPCR; computational methods; oligomerization