Washida N, Kitawaki J, Higashiyama T, Matsui S, Osawa Y
Endocrine Biochemistry Department, Hauptman-Woodward Medical Research Institute, Inc., Buffalo, NY 14203-1196, USA.
Steroids. 1996 Mar;61(3):126-32. doi: 10.1016/0039-128x(95)00215-c.
We produced a murine monoclonal antibody (MAb) to human placental aromatase cytochrome P450. This MAb, designated MAb3-2C2, was selected on its ability to suppress aromatase activity. The specificity of this MAb was assessed by selective immunoprecipitation of 125I-labeled aromatase cytochrome P450 as well as by the identification of a 55-kDa protein, which was enriched and purified by immunoaffinity chromatography on a MAb-coupled Sepharose 4B column. The MAb was able to suppress both human placental and ovarian microsomal aromatase. Species differences of aromatase were recognized by MAb3-2C2 on the basis of differential immunosuppression of aromatase activity. The antibody had no effect on non-aromatase cytochrome P450s. MAb3-2C2 gave negative results with human placental aromatase P450 in the Western blot analysis. The data presented indicate that MAb3-2C2 is specific for aromatase cytochrome P450 and that its epitope is located in a fragile tertiary conformation of the enzyme, thus making it capable of sensitively affecting catalysis.
我们制备了一种针对人胎盘芳香化酶细胞色素P450的鼠单克隆抗体（MAb）。这种单克隆抗体命名为MAb3 - 2C2，是根据其抑制芳香化酶活性的能力筛选出来的。通过对125I标记的芳香化酶细胞色素P450进行选择性免疫沉淀以及鉴定一种55 kDa的蛋白质来评估该单克隆抗体的特异性，该蛋白质通过在与单克隆抗体偶联的琼脂糖4B柱上进行免疫亲和层析而得到富集和纯化。该单克隆抗体能够抑制人胎盘和卵巢微粒体中的芳香化酶。MAb3 - 2C2基于对芳香化酶活性的不同免疫抑制作用识别出了芳香化酶的物种差异。该抗体对非芳香化酶细胞色素P450没有影响。在蛋白质免疫印迹分析中，MAb3 - 2C2与人胎盘芳香化酶P450呈阴性结果。所呈现的数据表明，MAb3 - 2C2对芳香化酶细胞色素P450具有特异性，并且其表位位于该酶的一个脆弱的三级结构中，从而使其能够敏感地影响催化作用。
