A chemiluminescence-based assessment of androgen-binding activity in a large pedigree affected with androgen insensitivity syndrome.
A chemiluminescence-based assessment of androgen-binding activity in a large pedigree affected with androgen insensitivity syndrome.
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对受雄激素不敏感综合征影响的大型家系的雄激素结合活性进行基于化学发光的评估。
DOI：
10.1002/bio.973
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
Zewei Luo
中科院分区：
文献类型：
--
作者：
Jiu;Ming;Rongmei Zhang;Zhi;Jie Chen;Zewei Luo
我们报道了一种新的基于化学发光(CL)的方法来检测雄激素受体的配体结合活性。该方法的核心是在H(2)O(2)-微量过氧化物酶体系中用类固醇化学发光标记物代替传统的配体结合试验中的放射性标记物，通过光测量结合的化学发光标记物来测定结合活性。用7例正常男性生殖器皮肤成纤维细胞(GSF)对该方法的性质和可靠性进行了研究和验证。该方法精密度高(B(Max)和K(D)的变异系数均为7%)，在各Scatchard线性回归分析中具有较高的相关性(r>；0.93)。该方法只需使用放射性标记方法所需细胞的四分之一(约40000个细胞/数据点)即可确定雄激素结合特性。通过对来自一个雄激素不敏感综合征大家族的几个患者的GSFs的结合实验，说明了该方法的有效性。家系的显著特征是所有患者都有相同的雄激素受体Arg840Cys替换，但在男性性发育障碍中表现出高度的表型变异。为本研究选择的患者代表了这种表型变异的广泛范围。因此，这项研究为该突变的多效性效应提供了见解。总之，基于化学发光的方法可以作为一种有效、准确和可靠的方法来替代放射性标记方法，并且与相应的方法相比具有简单、经济、健康和环境安全的优点。
We report a novel chemiluminescence (CL)-based method for assaying the ligand-binding activity of the androgen receptor. The central parts of this method are the utilization of the steroid CL marker as the replacement of the radioactive label in the conventional ligand-binding assay and the determination of the binding activity by the light measurement of the bound CL-label under an H(2)O(2)-microperoxidase system. The properties and reliability of this assay were investigated and verified using genital skin fibroblasts (GSF) from seven normal males. The method is precise (CV < 7% for both B(max) and K(d)) with high correlation coefficients (r > 0.93) in each Scatchard linear regression analysis. This assay can determine the androgen binding properties using only a quarter of the cells (approximately 40 000 cells/data point) of that required by the radiolabelling approach. The utility of the method was illustrated by binding experiment on the GSFs of several patients from a large Chinese family affected with androgen insensitivity syndrome. The familial distinct feature is that all patients shared an identical Arg840Cys substitution in the androgen receptor but displayed high phenotypic variation in disorders of male sexual development. The patients selected for the present study represent a wide spectrum of this phenotypic variation. This study thus provides insights on the pleiotropic effects of the mutation. In conclusion, the CL-based method can serve as an effective, precise and reliable replacement for the radiolabelling approach and has the advantages of simplicity, cost-effectiveness and health and environmental safety over the counterpart.
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DOI：
10.1152/ajpcell.1982.243.1.c81
发表时间：
1982
期刊：
The American journal of physiology
影响因子：
--
作者：
Airhart,J;Arnold,JA;Stirewalt,WS;Low,RB
通讯作者：
Low,RB
DOI：
10.1210/jcem-55-3-465
发表时间：
1982-09
期刊：
The Journal of clinical endocrinology and metabolism
影响因子：
--
作者：
James E. Griffin;Joy L. Durrant
通讯作者：
James E. Griffin;Joy L. Durrant
影响因子：
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作者：
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通讯作者：
Zhong-xun Zhou;M. Lane;J. A. Kemppainen;F. S. French;E. M. Wilson
影响因子：
20.3
作者：
QUIGLEY, CA;DEBELLIS, A;FRENCH, FS
通讯作者：
FRENCH, FS
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DOI：
10.1172/jci117507
发表时间：
1994
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Marcelli,M;Zoppi,S;Wilson,CM;Griffin,JE;McPhaul,MJ
通讯作者：
McPhaul,MJ