Purvis K, Brekke I, Rui H, Sunde A
J Steroid Biochem. 1986 Dec;25(6):1023-5. doi: 10.1016/0022-4731(86)90339-0.
Kinetic constants for the 5 alpha-reductase were determined in freshly isolated epithelial cells from the rat ventral prostate. Studies were also performed on stromal tissue but not isolated stromal cells for comparison. Secretory and non-secretory epithelial cells were separated by centrifugation in a Percoll gradient. Both epithelial cell populations metabolized testosterone to predominantly 5 alpha-dihydrotestosterone (5 alpha-DHT), although when expressed per cell the capacity for conversion was 3-4-fold higher for secretory cells (7.4 pmol/min/10(6) cells) than for non-secretory cells (2.3 pmol/min/10(6) cells; P less than 0.01 in 4 separate studies). When compared per mg cytosol protein this difference became non-significant. Stromal tissue contained a 5 alpha-reductase Vmax (expressed) per mg protein) which was comparable to the non-secretory cell enzyme. Lineweaver-Burke plots revealed different Km values for the different cell populations (12.5, 5.9 and 4.7 microM for secretory, non-secretory and stromal cells, respectively) suggesting the presence of different isoforms of the enzyme, or differences in the intracellular concentrations of enzyme antagonists.
在从大鼠腹侧前列腺新鲜分离的上皮细胞中测定了5α-还原酶的动力学常数。还对基质组织进行了研究,但未对分离的基质细胞进行研究以作比较。通过在Percoll梯度中离心分离分泌性和非分泌性上皮细胞。两种上皮细胞群体都将睾酮主要代谢为5α-双氢睾酮(5α-DHT),尽管以每个细胞计算时,分泌细胞的转化能力(7.4 pmol/分钟/10⁶个细胞)比非分泌细胞(2.3 pmol/分钟/10⁶个细胞)高3 - 4倍(在4项独立研究中P小于0.01)。当以每毫克胞浆蛋白进行比较时,这种差异变得不显著。基质组织每毫克蛋白所含的5α-还原酶Vmax(表达量)与非分泌细胞的酶相当。Lineweaver-Burke图显示不同细胞群体的Km值不同(分泌细胞、非分泌细胞和基质细胞分别为12.5、5.9和4.7 microM),这表明存在该酶的不同同工型,或者酶拮抗剂的细胞内浓度存在差异。