Detection of isoflavones and phytoestrogen-rich plant extracts binding to estrogen receptor β using a yeast-based fluorescent assay.

Sofija Bekić, Edward Petri, Sanja Krstić, Andjelka Ćelić, Suzana Jovanović-Šanta

Journal: Analytical biochemistry 2025;690():115529

PMID: 38582243

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Abstract

Alchemilla vulgaris L., Trifolium pratense L. and Glycyrrhiza glabra L. are important remedies in traditional medicine, known for many usages, including treating gynecological diseases. Despite folkloric use of the plant materials, there is a lack of scientific data to support their therapeutic application. The aims of the present study were to evaluate the relative binding affinities (RBAs) of plant-derived phytoestrogens for estrogen receptor β (ERβ) using fluorescent biosensor in yeast and to apply this assay for the assessment of the potential of plant materials towards ERs and treatment of estrogen-related disorders. Ligand-binding domain of ERβ fused with yellow fluorescent protein (ERβ LBD-YFP) was expressed in S. cerevisiae and fluorescence was detected by fluorimetry and fluorescence microscopy. Structural basis for experimental results was explored by molecular docking. Yeast-based fluorescent assay was successfully optimized and applied for identification of natural phenolic compounds and phytoestrogen-rich plant extracts that interact with ERβ-LBD, making this biosensor a valuable tool for screening estrogenic potential of a variety of plant extracts. This assay can be used for preliminary testing of plant-derived or fungal extracts, but also other sources of environmental substances with ER-modulating activity in order to assess their possible effects on the female reproductive system.

Address: Department of Chemistry, Biochemistry and Environmental Protection, Faculty of Sciences University of Novi Sad, Trg Dositeja Obradovića 3, 21000, Novi Sad, Serbia. Electronic address: [email protected].; Department of Biology and Ecology, Faculty of Sciences University of Novi Sad, Trg Dositeja Obradovića 2, 21000, Novi Sad, Serbia. Electronic address: [email protected].; Department of Chemistry, Biochemistry and Environmental Protection, Faculty of Sciences University of Novi Sad, Trg Dositeja Obradovića 3, 21000, Novi Sad, Serbia; Institute of Pharmaceutical Sciences, University of Graz, Beethovenstraße 8, 8010, Graz, Austria. Electronic address: [email protected].; Department of Biology and Ecology, Faculty of Sciences University of Novi Sad, Trg Dositeja Obradovića 2, 21000, Novi Sad, Serbia. Electronic address: [email protected].; Department of Chemistry, Biochemistry and Environmental Protection, Faculty of Sciences University of Novi Sad, Trg Dositeja Obradovića 3, 21000, Novi Sad, Serbia. Electronic address: [email protected].

DOI: 10.1016/j.ab.2024.115529

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MeSH Terms: Estrogen Receptor beta, Phytoestrogens, Plant Extracts, Saccharomyces cerevisiae, Isoflavones, Molecular Docking Simulation, Biosensing Techniques, Humans, Trifolium, Glycyrrhiza