Estrogen receptor stereochemistry: Receptor binding and hormonal responses

Abstract

Estrogen stimulation of the uterus elicits a spectrum of biochemical responses which are customarily linked together. DES and certain structural analogs, indenestrol A (IA), indenestrol B (IB), indanestrol (I), and pseudo DES (PD), were used as probes to segregate various genomic responses previously considered interrelated, most notably the events of specific protein synthesis, DNA synthesis, and mitosis. These compounds have poor uterotrophic activity; except for I, they interact specifically with mouse uterine estrogen receptors (ER) with high affinity. All translocate stoichiometrically similar amounts of ER complex to the nucleus. IA and IB possess a single chiral carbon atom and exist as a mixture of enantiomers (ENT). We investigated whether the poor biological activity of IA could be explained by differential activity of the enantiomers. The IA ENT were separated to > 98% purity using a chirally active HPLC column. Competitive binding assays to cytosolic ER demonstrated a stereochemical chiral preference. This preference was also evident from nuclear ER translocation experiments. IB was as active as DES to induce mouse uterine glucose 6-phosphate dehydrogenase (G-6-PD), while the other compounds had weak activity. Induction of cytosolic progesterone receptor (PR) was stimulated by all the DES compounds. Ornithine decarboxylase (ODC) was stimulated 600% by DES and 180% by IB; the other compounds had no significant activity. Uterine DNA synthesis was increased by DES and IB. Thymidine autoradiography indicated nuclear labeling was occurring primarily in luminal epithelium. Treatment with PD increased uterine cell height but not cell numbers, suggesting the two responses are not necessarily interdependent as previously thought and may require two separate receptor interactions. Such a probe should be useful in studying the individual events involved in estrogen-induced uterine growth. These data also indicate that induction of ER, PR and G-6-PD are not coupled. Therefore, stimulation of a certain uterine response may depend on the structure of the particular ligand receptor complex formed, and its interaction may be regulated by specificity at the genomic acceptor site.