Infoscience

Journal article

Diffusion-Time Distribution Analysis Reveals Characteristic Ligand-Dependent Interaction Patterns of Nuclear Receptors in Living Cells

Nuclear receptors initiate transcription, interact with regulatory proteins, and are influenced by hormones, drugs, and pollutants. Herein, we discover ligand-specific mobility patterns of human estrogen receptor-a (ER) in living cells using diffusion-time distribution anal. (DDA). This novel method, based on fluorescence correlation spectroscopy (FCS), is esp. suited to unraveling multiple protein interactions in vivo at native expression levels. We found that ER forms a limited no. of distinct complexes with a varying population by dynamic interaction with other nuclear components. Dose-response curves of different ligands could be obtained for each receptor interaction. The potential to identify interacting proteins was demonstrated by comparing DDA of the ER cofactor SRC-3 attached to yellow fluorescent protein (YFP) with those of YFP-ER. Our findings open up new routes to elucidating transcription regulation and to detecting and distinguishing pharmacol. and toxicol. active compds. in vivo. Moreover, DDA provides a general approach to monitoring biochem. networks in individual living cells. [on SciFinder (R)]

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