Bioluminescent Indicator for Highly Sensitive Analysis of Estrogenic Activity in a Cell-Based Format

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• Estrogens are critical hormones regulating diverse physiological processes.
• Accurate quantification of nanomolar estrogens in live cells is challenging but vital for drug discovery and medical diagnostics.
• Existing methods lack the sensitivity and speed required for high-throughput screening.

Purpose of the Study:

• To develop a sensitive and quantitative bioluminescent indicator for detecting low concentrations of estrogens.
• To enable rapid screening of estrogenic compounds and potential therapeutic inhibitors.

Main Methods:

• Constructed a bioluminescent indicator using a ligand-binding domain of an estrogen receptor fused to a binding peptide.
• Integrated the receptor-peptide construct between split fragments of a luciferase mutant.
• Assessed the indicator's response to 17β-estradiol and other estrogenic compounds.

Main Results:

• The indicator demonstrated restored bioluminescence upon binding to 17β-estradiol at concentrations above 1.0 × 10^-10 M.
• Achieved a high signal-to-background ratio for quantitative estrogen detection.
• The indicator responded to estrogen agonists but not to antagonists.

Conclusions:

• The developed bioluminescent indicator provides a sensitive and quantitative method for detecting low-concentration estrogens.
• This tool is suitable for rapid, high-throughput screening of estrogenic compounds and inhibitors.
• Facilitates accelerated drug discovery and development in areas related to estrogenic pathways.