Competitive Binding Assay for the G-Protein-Coupled Receptor 30 (GPR30) or G-Protein-Coupled Estrogen Receptor (GPER)

  • Thomas Thekkumkara
  • Russell Snyder
  • Vardan T. Karamyan
Part of the Methods in Molecular Biology book series (MIMB, volume 1366)


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.

Key words

2-Methoxyestradiol GPR30 G-protein-coupled receptor 30 GPER G-protein-coupled estrogen receptor Endoplasmic reticulum membrane Radioligand binding Competitive binding 



The authors would like to acknowledge the laboratory personnel responsible for optimization of this protocol; in particular, the authors wish to acknowledge the graduate student Siva Koganti and fellow Upendra Gumaste. Studies were funded by NIH Grant #DK072140.


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Thomas Thekkumkara
    • 1
  • Russell Snyder
    • 1
  • Vardan T. Karamyan
    • 2
  1. 1.Department of Biomedical SciencesTexas Tech University Health Sciences CenterAmarilloUSA
  2. 2.Department of Pharmaceutical SciencesTexas Tech University Health Sciences CenterAmarilloUSA

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