Namkung P C, Stanczyk F Z, Cook M J, Novy M J, Petra P H
Department of Ob/Gyn, University of Washington, Seattle 98195.
J Steroid Biochem. 1989 May;32(5):675-80. doi: 10.1016/0022-4731(89)90511-6.
We have recently shown that the metabolic clearance of testosterone in plasma is directly dependent on sex steroid-binding protein (SBP or SHBG) levels [J. steroid. Biochem. 22, 739 (1986)]. In order to further understand the relationship between these two parameters, we have measured the half-life of SBP in plasma of female rhesus monkeys. SBP was purified to homogeneity from pooled Macaca nemestrina serum, and iodinated with 125I. The labeled protein ([125I]nSBP) was purified by chromatography on DEAE-agarose and fractions identified as immunologically reactive against anti-human SBP were collected. Protein purity of [125I]nSBP was established by SDS gel electrophoresis using an unlabelled Macaca nemestrina SBP as standard. The labeled protein was infused intravenously into two different adult female Macaca mulatta (rhesus) monkeys. Plasma samples were collected at short intervals during the first 24 h after infusion, and then daily for 7-9 days. The clearance profile of labeled SBP in plasma was quantitated by radioactivity measurement and immunoprecipitation. Analysis of the results indicate that the rate of SBP clearance in plasma has two components, the t1/2 (app) of the first component is 7.5 h (r = 0.94), and the t1/2 (app) of the second component is 3.95 days (r = 0.95). Over 90% of the injected 125I-nSBP was removed from plasma within 24 h at a rate corresponding to the t1/2 (app) of the first component. The data indicate that most of the SBP rapidly distributes into extravascular spaces during the first 24 h following infusion, and are consistent with the hypothesis that SBP may be directly involved in sex steroid hormone transport into tissues.
我们最近发现,血浆中睾酮的代谢清除率直接取决于性类固醇结合蛋白(SBP或SHBG)水平[《类固醇生物化学杂志》22, 739 (1986)]。为了进一步了解这两个参数之间的关系,我们测量了雌性恒河猴血浆中SBP的半衰期。从混合的食蟹猴血清中纯化SBP至同质,并使用125I进行碘化。标记的蛋白质([125I]nSBP)通过DEAE-琼脂糖柱层析纯化,收集与抗人SBP具有免疫反应性的级分。以未标记的食蟹猴SBP为标准,通过SDS凝胶电泳确定[125I]nSBP的蛋白质纯度。将标记的蛋白质静脉注射到两只不同的成年雌性恒河猴体内。在注射后的头24小时内每隔短时间收集血浆样本,然后每天收集7 - 9天。通过放射性测量和免疫沉淀对血浆中标记SBP的清除情况进行定量分析。结果分析表明,血浆中SBP的清除率有两个成分,第一个成分的t1/2(表观)为7.5小时(r = 0.94),第二个成分的t1/2(表观)为3.95天(r = 0.95)。超过90%的注射125I-nSBP在24小时内以与第一个成分的t1/2(表观)相对应的速率从血浆中清除。数据表明,大多数SBP在注射后的头24小时内迅速分布到血管外间隙,这与SBP可能直接参与性类固醇激素转运到组织中的假设一致。