Inhibition of nucleic acid and chromatin binding of the rat prostate androgen receptor by pyridoxal phosphate, heparin and cibacron blue

Abstract

The androgen receptor from rat prostate binds 5α-dihydro-testosterone and related androgenic steroids at a steroid binding site and in addition shows selective binding to structures related to nucleic acids (chromatin binding site). Cytoplasmic androgen receptor, labeled with the synthetic androgen methyltrienolone (R 1881) was readily bound at 4°C by 2',5'-ADP-agarose, DNA-cellulose and phosphocellulose. The binding to ADP-agarose and DNA-cellulose was used as a model for study of the nucleic acid binding site of the receptor. Complete elution of androgen receptors from these matrices could be obtained with low concentrations of pyridoxal phosphate (10 mM), heparin (0.2 mg/ml) and Cibacron blue (0.4 mM). Sodium molybdate (10 mM) did not interfere significantly with binding of the androgen receptor to ADP-agarose and had little effect on elution of receptors from the gel. Pyridoxal phosphate, heparin and Cibacron blue in low ionic strength buffers were also shown to be very effective for the extraction of androgen receptors from nuclei of rat prostatic tissue. These results suggest gross similarities in the structure of the androgen receptor with activated forms of receptors for corticoids, estrogens and progestins with respect to nucleic acid binding.