Matsubayashi A
Kanagawa Shigaku. 1989 Jun;24(1):38-58.
The low molecular weight inhibitor (LMWI) and the translocation of dihydrotestosterone (DHT)-androgen binding protein (ABP) complex of the cytosol to the nuclei in rat submandibular gland (SMG) was studied by dialysis, ultrafiltration and glycerol linear gradient centrifugation procedures. Prebound cytosol was obtained by the incubation with 3H-DHT at 4 degrees C for 3hr in the presence or absence of 100 fold excess of unlabeled DHT prior to contact to nuclei. When prebound cytosol was dialyzed or ultrafiltrated, the binding ability of 3H-DHT-ABP complex to nuclei was increased up to 3 times of the control (nontreated prebound cytosol). The sedimentation rate of 3H-DHT-ABP complex by glycerol linear gradient centrifugation was 4S for dialyzed or ultrafiltrated prebound cytosol and 8S for control. These transformation of 8S to 4S and activation to the nuclear binding of 3H-DHT-ABP complex by dialysis or ultrafiltration were inhibited by molybdate in the prebound medium. The similar transformation of 8S to 4S and activation to the nuclear binding of 3H-DHT-ABP complex was shown in heated prebound cytosol. These results indicate that the LMWI regulate activation of 3H-DHT-ABP complex. The molecular weight of the dialyzed LMWI were estimated as 7,000 by dialysis or ultrafiltration membrane and SDS-PAGE. From the in vitro two step binding assay, it was revealed that rat SMG contained the low molecular weight protein (M. W. 7,000) in cytosol having a inhibitory effect on the translocation to the nuclei of 3H-DHT-ABP complex.
采用透析、超滤和甘油线性梯度离心法，研究了大鼠下颌下腺（SMG）中低分子量抑制剂（LMWI）以及胞质溶胶中双氢睾酮（DHT）-雄激素结合蛋白（ABP）复合物向细胞核的转运。在与细胞核接触之前，通过在4℃下用3H-DHT孵育3小时，在存在或不存在100倍过量未标记DHT的情况下获得预结合的胞质溶胶。当对预结合的胞质溶胶进行透析或超滤时，3H-DHT-ABP复合物与细胞核的结合能力增加至对照（未处理的预结合胞质溶胶）的3倍。通过甘油线性梯度离心法，透析或超滤后的预结合胞质溶胶中3H-DHT-ABP复合物的沉降速率为4S，对照为8S。预结合介质中的钼酸盐可抑制透析或超滤导致的8S向4S的转变以及3H-DHT-ABP复合物与细胞核结合的激活。加热后的预结合胞质溶胶中也显示出8S向4S的类似转变以及3H-DHT-ABP复合物与细胞核结合的激活。这些结果表明，LMWI调节3H-DHT-ABP复合物的激活。通过透析或超滤膜以及SDS-PAGE估计，透析后的LMWI分子量为7000。从体外两步结合试验可知，大鼠SMG的胞质溶胶中含有低分子量蛋白（分子量7000），对3H-DHT-ABP复合物向细胞核的转运具有抑制作用。
