Identification of a polymorphically expressed member of the human cytochrome P-450III family.
Identification of a polymorphically expressed member of the human cytochrome P-450III family.
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发表时间：
1989-07
影响因子：
3.6
通讯作者：
S. Wrighton;B. Ring;P. Watkins;M. Vandenbranden
中科院分区：
文献类型：
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作者：
S. Wrighton;B. Ring;P. Watkins;M. Vandenbranden
人肝细胞色素 P-450III 家族包含两种高度相关的细胞色素 P-450 (P450) 形式：HLp 和 P450NF（在成人中表达）和一种形式 HLp2（在胎儿中表达）。使用抗 HLp 抗体对 46 个肝脏样本进行免疫印迹分析，结果表明，除了 HLp 之外，11 个样本还含有一种以前未检测到的较高分子量蛋白（称为 HLp3）。 HLp3的表达与患者的年龄、性别、吸烟习惯或用药史无关。该蛋白经纯化后发现为P450，分子量为52,000。 Ouchterlony 使用多克隆抗 HLp 抗体进行分析，得出 HLp3 与 HLp 和 HLp2 之间的部分同一性系。此外，这三种蛋白质之间的结构比较，包括氨基末端氨基酸分析和肽图分析，表明HLp3、HLp2和HLp是高度相关但不同的蛋白质。在重组系统中，HLp 和 HLp3 被发现可在 6 β 和 2 β 位点羟基化睾酮。在微粒体中，发现睾酮在 2β、6α、6β、15β 和 18 位点的羟基化率以及未知物的形成与总 HLp 相关蛋白的水平相关，并且这些活性被抗 HLp 抗体抑制。总之，我们的数据表明 HLp3 是人类 P450III 家族的成员，并且是多态性表达的。
The human liver cytochrome P-450III family contains two highly related forms of cytochrome P-450 (P450), HLp and P450NF, that are expressed in the adult and a form, HLp2, that is expressed in the fetus. Immunoblot analyses of 46 liver specimens developed with an anti-HLp antibody demonstrated that, in addition to HLp, 11 specimens contained a previously undected higher molecular weight protein (termed HLp3). The expression of HLp3 did not correlate with the age, gender, smoking habits, or drug history of the patients. This protein was purified and found to be a P450 with a molecular weight of 52,000. Ouchterlony analyses using a polyclonal anti-HLp antibody yielded lines of partial identity between HLp3 and both HLp and HLp2. In addition, structural comparisons between these three proteins, including amino-terminal amino acid analyses and peptide mapping, indicated that HLp3, HLp2, and HLp are highly related but distinct proteins. In reconstituted systems, HLp and HLp3 were found to hydroxylate testosterone at the 6 beta and 2 beta positions. In microsomes, the rates of hydroxylation of testosterone at the 2 beta, 6 alpha, 6 beta, 15 beta, and 18 positions and the formation of an unknown were found to correlate with the levels of total HLp-related protein and these activities were inhibited by anti-HLp antibodies. In conclusion, our data demonstrate that HLp3 is a member of the human P450III family and is polymorphically expressed.