Expression of a full-length cDNA encoding bovine adrenal cytochrome P450C21.
Expression of a full-length cDNA encoding bovine adrenal cytochrome P450C21.
复制标题
编码牛肾上腺细胞色素 P450C21 的全长 cDNA 的表达。
DOI：
10.1016/0003-9861(89)90164-1
复制
发表时间：
1989
影响因子：
3.9
通讯作者：
Waterman,MR
中科院分区：
文献类型：
--
作者：
Lorence,MC;Trant,JM;Mason,JI;Bhasker,CR;Fujii-Kuriyama,Y;Estabrook,RW;Waterman,MR
用已知结构的部分cDNA构建了牛肾上腺皮质P450 C21的两个全长cDNA克隆。在COS-1细胞中表达后，测定P450 C21的底物特异性。在检测的18种类固醇中，发现孕酮、17α-羟孕酮和11β，17 α-二羟孕酮是唯一可作为该肾上腺酶底物的类固醇，其底物特异性程度远高于已报道的肝21-羟化酶。17α-羟孕酮的Vmax是孕酮的2.5倍，而Δ5-甾体不能作为该酶的底物。两种cDNA之间的差异位于氨基酸401处，其中一种所得酶含有酪氨酸，而另一种含有组氨酸。这种氨基酸差异似乎对肾上腺P450 C21的动力学特性没有影响。
Two full-length cDNA clones encoding bovine adrenocortical P450C21have been constructed in a eukaryotic expression vector using partial-length cDNAs whose structures have been previously reported. Following expression of these cDNAs in COS 1 cells, the substrate specificity of P450C21was determined. Of the 18 steroids tested, progesterone, 17α-hydroxyprogesterone, and 11β,17α-dihydroxyprogesterone were found to be the only steroids to serve as substrates for this adrenal enzyme, a much higher degree of substrate specificity than has been reported for a hepatic 21-hydroxylase. TheVmaxfor 17α-hydroxyprogesterone was 2.5 times greater than that for progesterone, whereas Δ5-steroids were unable to serve as substrate for this enzyme. A difference between the two cDNAs is located at amino acid 401 where one resultant enzyme contains tyrosine while the other contains histidine. This amino acid difference appears to have no effect on the kinetic properties of adrenal P450C21