title: Competitive Binding Assay for the G-Protein-Coupled Receptor 30 (GPR30) or G-Protein-Coupled Estrogen Receptor (GPER).
abstract: The role of 2-methoxyestradiol is becoming a major area of investigation because of its therapeutic utility, though its mechanism is not fully explored. Recent studies have identified the G-protein-coupled receptor 30 (GPR30, GPER) as a high-affinity membrane receptor for 2-methoxyestradiol. However, studies aimed at establishing the binding affinities of steroid compounds for specific targets are difficult, as the tracers are highly lipophilic and often result in nonspecific binding in lipid-rich membrane preparations with low-level target receptor expression. 2-Methoxyestradiol binding studies are essential to elucidate the underlying effects of this novel estrogen metabolite and to validate its targets; therefore, this competitive receptor-binding assay protocol was developed in order to assess the membrane receptor binding and affinity of 2-methyoxyestradiol.
authors.affiliation: Department of Biomedical Sciences, Texas Tech University Health Sciences Center, 1300 Coulter Drive, Amarillo, TX, 79106, USA. thomas.thekkumkara@ttuhsc.edu.
authors.forename: Thomas
authors.initials: T
authors.lastname: Thekkumkara
authors.affiliation: Department of Biomedical Sciences, Texas Tech University Health Sciences Center, 1300 Coulter Drive, Amarillo, TX, 79106, USA.
authors.forename: Russell
authors.initials: R
authors.lastname: Snyder
authors.affiliation: Department of Pharmaceutical Sciences, Texas Tech University Health Sciences Center, 1300 South Coulter, Amarillo, TX, 79106, USA.
authors.forename: Vardan T
authors.initials: VT
authors.lastname: Karamyan
mesh_terms: D000077584:2-Methoxyestradiol; D000818:Animals; D052117:Benzodioxoles / Q000378:metabolism; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004958:Estradiol / Q000031:analogs & derivatives* / Q000378:metabolism; D008024:Ligands; D011485:Protein Binding; D011804:Quinolines / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D057188:Workflow
chemical_list: C568862:4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta(c)quinoline; D052117:Benzodioxoles; C520305:Gper1 protein, rat; D008024:Ligands; D011804:Quinolines; D043562:Receptors, G-Protein-Coupled; D004958:Estradiol; D000077584:2-Methoxyestradiol
keywords: 2-Methoxyestradiol; Competitive bind ing; Endoplasmic reticulum membrane; G-protein-coupled estrogen receptor; G-protein-coupled receptor 30; GPER; GPR30; Radioligand bind ing