Characterization of two nuclear androgen receptors in Atlantic croaker: comparison of their biochemical properties and binding specificities.

Two distinct androgen receptors (ARs) with different characteristics were identified in the brain and ovary of Atlantic croaker, Micropogonias undulatus. A nuclear AR, AR1, was identified in the brain that had high affinity binding sites for testosterone (T; Kd, 1.1 +/- 0.15 nM; binding capacity, 1.4 +/- 0.14 pmol/g tissue; n = 16). A second nuclear AR, AR2, was found in the ovary that had high affinity binding sites for 5alpha-dihydrotestosterone (DHT; Kd, 0.62 +/- 0.1 nM; binding capacity, 0.38 +/- 0.06 pmol/g tissue; n = 14). AR2 has physiochemical properties similar to those of other vertebrate ARs. AR2 has high affinity binding for a broad spectrum of natural and synthetic androgens, including 17alpha-methyl-5alpha-dihydrotestosterone, which has a relative binding affinity of DHT = 100% > T > mibolerone > 11-ketotestosterone = 16%, a rapid association (t1/2, 44 min) and a slow dissociation (t1/2, 45 h) rate, as well as specific binding to purified DNA. The cytosolic AR2 interacts with heat shock proteins in a manner similar to other steroid receptors, as sodium molybdate stabilizes the receptor, and it has a 7.4-7.8S sedimentation coefficient in a 5-20% sucrose gradient. In contrast, AR1 is highly specific for only a few androgens, with T = 100% relative binding affinity >> DHT >> 11-ketotestosterone > mibolerone > 17alpha-methyl-5alpha-dihydrotestosterone = 0, has rapid association (t1/2, 15 min) and dissociation (t1/2, 2.6 +/- 0.7 h) rates, and has specific binding to purified DNA upon heat activation. The cytosolic binding component sediments at 5.6-5.7S in a 5-20% sucrose gradient and is not affected by sodium molybdate, which suggests that AR1 does not interact with heat shock proteins in the usual manner. This is the first report of the presence of two different nuclear ARs displaying markedly different steroid binding specificities within a single vertebrate species.