ISOLATION OF A FULL-LENGTH CDNA INSERT ENCODING HUMAN AROMATASE SYSTEM CYTOCHROME-P-450 AND ITS EXPRESSION IN NONSTEROIDOGENIC CELLS
ISOLATION OF A FULL-LENGTH CDNA INSERT ENCODING HUMAN AROMATASE SYSTEM CYTOCHROME-P-450 AND ITS EXPRESSION IN NONSTEROIDOGENIC CELLS
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DOI：
10.1073/pnas.85.23.8948
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发表时间：
1988-12-01
影响因子：
11.1
通讯作者：
SIMPSON, ER
中科院分区：
文献类型：
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作者：
CORBIN, CJ;GRAHAMLORENCE, S;SIMPSON, ER
本文报道了一个与编码人芳香化酶系统细胞色素P-450的mRNA互补的全长cDNA插入片段的分离和克隆。插入片段含有编码503个氨基酸的蛋白质的开放阅读框。该基因显然是细胞色素P-450基因超家族的成员，因为该序列包含与其他成员具有显著同源性的区域，特别是推定的跨膜区域、I螺旋、Ozols和血红素结合区域。将该cDNA插入到经修饰的pCMV载体中，并在COS-1猴肾肿瘤细胞中表达。所表达的蛋白质在大小上与人胎盘芳香化酶系统细胞色素P-450相似，如通过免疫印迹分析检测到的，并且催化雄烯二酮、睾酮和16 α-雌二醇的芳构化。羟基雄烯二酮这种活性被已知的芳香酶抑制剂4-羟基雄烯二酮和益康唑抑制。因此，芳构化反应中涉及的几个步骤似乎是由一个单一的多肽链催化的，它可以代谢三种主要的生理底物。
The isolation and cloning of a full-length cDNA insert complementary to mRNA encoding human aromatase system cytochrome P-450 is reported. The insert contains an open reading frame encoding a protein of 503 amino acids. This gene is clearly a member of the cytochrome P-450 gene superfamily, because the sequence contains regions of marked homology to those of other members, notably a putative membrane-spanning region, I helix, Ozols, and heme-binding regions. The cDNA was inserted into a modified pCMV vector and expressed in COS-1 monkey kidney tumor cells. The expressed protein was similar in size to human placental aromatase system cytochrome P-450, as detected by immunoblot analysis and catalyzed the aromatization of androstenedione, testosterone, and 16.alpha.-hydroxyandrostenedione. This activity was inhibited by the known aromatase inhibitors, 4-hydroxyandrostenedione and econazole. Thus the several steps involved in the aromatization reaction appear to be catalyzed by a single polypeptide chain, which can metabolize the three major physiological substrates.