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人精液中雄激素结合蛋白结合能力的测定方法(作者译)

[The method for the measurement of the binding capacity of androgen binding protein in human semen (author's transl)].

作者信息

Oda T

出版信息

Nihon Sanka Fujinka Gakkai Zasshi. 1982 Jan;34(1):1-10.

PMID:7199550
Abstract

In the present study, the androgen-binding properties of human semen was evaluated by experimenting with incubation and the technique for separating bound from unbound hormones. Some of the properties of a protein in human seminal plasma (SPABP) that has a high affinity and low capacity for binding dihydrotestosterone (DHT) was found and studied. The dissociation constant (Kd) was 1.63 X 10(-9) M and the number of binding sites of 275.3 X 10(-12)M was obtained. The intraassay and interassay coefficient of variation of this assay method was less than 5.6% and 8.4%, respectively. DHT was displaced from the binding protein by testosterone (81%), estradiol (75%) and levonorgestrel (63%). The endogenous level of steroids in human seminal plasma did not interfere with the estimation of DHT binding sites. In order to investigate the binding of DHT to human spermatozoa, the separation of sperm from the incubation mixture was accomplished by low-field centrifugation followed by 3,000 X G for 10 minutes. Using this separation method, the optimal steady-state condition for DHT-binding was found to be an incubation time of one hour. The binding of DHT to spermatozoa was nonspecific in nature, that is, of high capacity and low affinity (Kd greater than 10(-6)M).

摘要

在本研究中,通过孵育实验以及分离结合态与非结合态激素的技术,对人类精液的雄激素结合特性进行了评估。发现并研究了人类精浆中一种对双氢睾酮(DHT)具有高亲和力和低结合容量的蛋白质(SPABP)的一些特性。解离常数(Kd)为1.63×10⁻⁹M,结合位点数量为275.3×10⁻¹²M。该检测方法的批内和批间变异系数分别小于5.6%和8.4%。睾酮(81%)、雌二醇(75%)和左炔诺孕酮(63%)可使DHT从结合蛋白上解离下来。人类精浆中类固醇的内源性水平不干扰DHT结合位点的测定。为了研究DHT与人类精子的结合,通过低场离心,然后在3000×G下离心10分钟,将精子从孵育混合物中分离出来。使用这种分离方法,发现DHT结合的最佳稳态条件是孵育时间为1小时。DHT与精子的结合本质上是非特异性的,即具有高容量和低亲和力(Kd大于10⁻⁶M)。

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