ESTROGEN SYNTHETASE: CHARACTERIZATION AND CONTROL

我们建议从人胎盘中纯化细胞色素P-450 催化雄激素芳构化， 雌激素类。 酶的特征在于 底物特异性和动力学、理化性质和 部分氨基酸序列。 我们还建议， 纯化蛋白的多克隆和单克隆抗体。 此外，我们建议研究几个 作为结构探针的活性位点定向抑制剂的类型 酶的功能。 两种可逆抑制剂 模拟雄激素底物的非甾体化合物， 一种新型的双功能类固醇抑制剂-将用于 绘制活性部位的几何形状。 几种化学 反应性类固醇--直接烷基化剂、自杀底物和 光亲和试剂-将用于共价标记 为了鉴定蛋白质序列中 包括活性位点。 这些抑制剂将具有启发性的用途 用于研究芳构化的分子事件的细节 反应并将在药物设计中应用 雌激素依赖性癌症的治疗。

We proposed to purify from human placenta the cytochrome P-450 enzyme which catalyzes the aromatization of androgens to estrogens. The enzyme will be characterized with respect to substate specificity and kinetics, physicochemical properties and partial amino acid sequence. We also proposed to prepare both polyclonal and monoclonal antibodies to the purified protein. In addition, we propose to investigate the interaction of several types of active site-directed inhibitors as probes of the structure and function of the enzyme. Two types of reversible inhibitors -- non-steroidal compounds which mimic the androgen substrates and a new type of bifunctional steroid inhibitor - will be used for mapping the active site geometry. Several types of chemically reactive steroids -- direct alkylators, suicide substrates and photo-affinity reagents - will be used to covalently label the protein in order to identify regions of the protein sequence which comprise the active site. The inhibitors will be of heuristic use for studying details of the molecular events of the aromatization reaction and will have an application in the design of drugs for the treatment of estrogen-dependent cancers.