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[人酸性附睾糖蛋白样分子的分析:cDNA的分离及组织定位]

[Analysis of the human acidic epididymal glycoprotein-like molecule: isolation of cDNA and tissue localization].

作者信息

Hayashi M

机构信息

Department of Obstetrics and Gynecology, Hokkaido University School of Medicine, Sapporo, Japan.

出版信息

Hokkaido Igaku Zasshi. 1995 Sep;70(5):743-53.

PMID:8543280
Abstract

Acidic epididymal glycoprotein (AEG) is an androgen-dependent, epididymal secretory protein assumed to play a major role in sperm maturation. In the present study, we isolated cDNA clones encoding the human AEG-like molecule and determined their nucleotide sequences. The deduced human AEG-like molecule was made up of 230 amino acids, excluding a signal peptide, and contained one potential N-linked glycosylation site. All cysteinyl residues were conserved between the human AEG-like molecule and the AEG molecules of rats and mice. The human AEG-like molecule was equally similar to the AEG molecules of rats and mice and a related testis-specific protein known as TPX1 of human and mice (approximately 40% amino acid sequence similarity). Northern blot analysis showed that the human AEG-like gene is expressed specifically in the epididymis. To identify the product of the human AEG-like gene, polyclonal antibody was produced by immunizing rabbits with a recombinant human AEG-like protein expressed in E. coli. This antibody detected a major band of 30 kD and a minor band of 26 kD in the caput, corpus, and cauda regions of the epididymis, the ductus deferens, the sperm, and the seminal plasma. Immunohistochemical analysis showed that the human AEG-like molecule is located in the lumen and epithelium of distal ductus efferentes and epididymal ducts, and on the postacrosomal region of the sperm head.

摘要

酸性附睾糖蛋白(AEG)是一种雄激素依赖性的附睾分泌蛋白,被认为在精子成熟过程中起主要作用。在本研究中,我们分离了编码人AEG样分子的cDNA克隆并确定了它们的核苷酸序列。推导的人AEG样分子由230个氨基酸组成,不包括信号肽,并且含有一个潜在的N-连接糖基化位点。人AEG样分子与大鼠和小鼠的AEG分子之间所有的半胱氨酸残基都是保守的。人AEG样分子与大鼠和小鼠的AEG分子以及一种相关的睾丸特异性蛋白(人及小鼠中称为TPX1)具有同等相似性(氨基酸序列相似性约为40%)。Northern印迹分析表明,人AEG样基因在附睾中特异性表达。为了鉴定人AEG样基因的产物,用在大肠杆菌中表达的重组人AEG样蛋白免疫兔子制备了多克隆抗体。该抗体在附睾的头段、体段和尾段区域、输精管、精子和精浆中检测到一条30 kD的主要条带和一条26 kD的次要条带。免疫组织化学分析表明,人AEG样分子位于输出小管远端和附睾管的管腔和上皮中,以及精子头部的顶体后区域。

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[Analysis of the human acidic epididymal glycoprotein-like molecule: isolation of cDNA and tissue localization].[人酸性附睾糖蛋白样分子的分析:cDNA的分离及组织定位]
Hokkaido Igaku Zasshi. 1995 Sep;70(5):743-53.
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