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.
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Acknowledgments
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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Thekkumkara, T., Snyder, R., Karamyan, V.T. (2016). Competitive Binding Assay for the G-Protein-Coupled Receptor 30 (GPR30) or G-Protein-Coupled Estrogen Receptor (GPER). In: Eyster, K.M. (eds) Estrogen Receptors. Methods in Molecular Biology, vol 1366. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3127-9_2
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DOI: https://doi.org/10.1007/978-1-4939-3127-9_2
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3126-2
Online ISBN: 978-1-4939-3127-9
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