CLONING AND EXPRESSION OF HUMAN LIVER DEHYDROEPIANDROSTERONE SULFOTRANSFERASE
CLONING AND EXPRESSION OF HUMAN LIVER DEHYDROEPIANDROSTERONE SULFOTRANSFERASE
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DOI：
10.1042/bj2890233
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发表时间：
1993-01-01
影响因子：
4.1
通讯作者：
FALANY, CN
中科院分区：
文献类型：
--
作者：
COMER, KA;FALANY, JL;FALANY, CN
脱氢表雄酮硫化转移酶(DHEA-ST)催化人体肝脏和肾上腺组织中多种类固醇的3‘-磷酸腺苷-5’-磷酸-硫酸盐硫化反应，是人体肝脏胆汁酸硫化的主要原因。本文报道了编码人肝DHEA-ST的基因的分离、鉴定和表达。DHEA-ST cDNA命名为DHEA-ST8，从Uni-Zap XR人肝脏cDNA文库中分离得到，全长1060个核苷酸，含有一个编码285-氨基酸的开放阅读框，其相对分子质量约为33 765 Da。体外翻译的DHEA-ST8产生了一个与DHEA-ST分子大小相同的蛋白质。在COS-7细胞中表达DHEA-ST8可产生一种具有活性的DHEA-ST蛋白，该蛋白与人肝DHEA-ST具有相同的分子质量，并可被兔抗(人)肝DHEA-ST抗体识别。对人类肝脏RNA的Northern-印迹分析检测到三个不同大小的转录本的存在；然而，对人类DNA的Southern-印迹分析表明，基因组中可能只有一个基因。这些结果描述了人类ST的克隆，它在人类肝脏和肾上腺中类固醇和胆汁酸的硫化过程中起着重要作用。
Dehydroepiandrosterone sulphotransferase (DHEA-ST) catalyses the 3'-phosphoadenosine 5'-phosphosulphate-dependent sulphation of a wide variety of steroids in human liver and adrenal tissue and is responsible for most, if not all, of the sulphation of bile acids in human liver. This report describes the isolation, characterization and expression of a cDNA which encodes human liver DHEA-ST. The DHEA-ST cDNA, designated DHEA-ST8, was isolated from a Uni-Zap XR human liver cDNA library and is composed of 1060 bp and contains an open reading frame encoding a 285-aminoacid protein with a molecular mass of approx, 33 765 Da. Translation of DHEA-ST8 in vitro generated a protein identical in molecular size with that of DHEA-ST. Expression of DHEA-ST8 in COS-7 cells produces an active DHEA-ST protein which is capable of sulphating DHEA, has the same molecular mass as human liver DHEA-ST and is recognized by rabbit anti(human) liver DHEA-ST) antibodies. Northern-blot analysis of human liver RNA detects the presence of three different size transcripts; however, Southern-blot analysis of human DNA suggests that only one gene may be present in the genome. These results describe the cloning of a human ST which has an important role in the sulphation of steroids and bile acids in human liver and adrenals.