Leenders F, Adamski J, Husen B, Thole H H, Jungblut P W
Max-Planck-Institut für experimentelle Endokrinologie, Hannover, Germany.
Eur J Biochem. 1994 May 15;222(1):221-7. doi: 10.1111/j.1432-1033.1994.tb18860.x.
We describe the cloning and sequencing of porcine 17 beta-estradiol dehydrogenase. The enzyme performs oxidation 360-fold more efficiently than reduction, both measured under optimal conditions. It is localized in specialized vesicles of epithelial cells. The cDNA clones were isolated from a lambda UNI ZAP XR library of porcine kidney and polymerase-chain-reaction-amplified from templates of uterus epithelium. In both tissues, the same enzyme is coded by a transcript of 2.9 kb. It contains a 69-b 5'-noncoding region, an open reading frame of 2211 b and a 3'-noncoding region of 624 b. The open reading frame of 737 amino acids with a predicted molecular mass 79,973 Da was confirmed by amino acid sequencing of peptides. The 80-kDa translation product is processed to the N-terminal 32-kDa enzyme, part of which is then covalently linked to actin. The estradiol dehydrogenase/actin complex and the 80-kDa translation product comigrate in SDS/PAGE.
我们描述了猪17β-雌二醇脱氢酶的克隆和测序。在最佳条件下测定，该酶的氧化效率比还原效率高360倍。它定位于上皮细胞的特殊小泡中。cDNA克隆是从猪肾的λUNI ZAP XR文库中分离出来的，并从子宫上皮模板进行聚合酶链反应扩增。在这两种组织中，同一种酶由一个2.9kb的转录本编码。它包含一个69bp的5'非编码区、一个2211bp的开放阅读框和一个624bp的3'非编码区。通过肽段的氨基酸测序证实了由737个氨基酸组成的开放阅读框，预测分子量为79973Da。80kDa的翻译产物被加工成N端32kDa的酶，其中一部分随后与肌动蛋白共价连接。雌二醇脱氢酶/肌动蛋白复合物和80kDa的翻译产物在SDS/PAGE中迁移率相同。
