ESTROGEN SYNTHETASE: CHARACTERIZATION AND CONTROL

雌激素合成酶：特征和控制

• 批准号: 3236533
• 负责人: LARRY E VICKERY
• 金额: $ 12.32万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-12-01 至 1990-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3236533
• 关键词: androgens; antibody formation; complementary DNA; cyclization; cytochrome P450; dihydrotestosterone; drug design /synthesis /production; endoplasmic reticulum; enzyme inhibitors; enzyme mechanism; estrogen inhibitor; estrogens; gel electrophoresis; hormone inhibitor; hormone regulation /control mechanism; hormone related neoplasm /cancer; laboratory mouse; ligase; molecular cloning; monoclonal antibody; placenta; protein sequence; radioimmunoassay; steroid hormone biosynthesis; taste

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.

我们建议从人胎盘中净化细胞色素P-450 将雄激素的芳香化催化为 雌激素。 酶将以相对于 取代特异性和动力学，物理化学特性和 部分氨基酸序列。 我们还建议准备两者 纯化蛋白质的多克隆和单克隆抗体。 此外，我们建议研究几种的相互作用 主动位置定向抑制剂的类型作为结构的探针 酶的功能。 两种可逆抑制剂 - 非甾体类化合物，它们模仿雄激素底物和 一种新型的双功能类固醇抑制剂 - 将用于 映射活动位点的几何形状。 化学上几种类型 反应性类固醇 - 直接烷基剂，自杀底物和 照片亲和力试剂 - 将用于共价标记 蛋白质以识别蛋白质序列的区域 包括活动站点。 抑制剂将是启发式的 用于研究芳香化分子事件的细节 反应，并将在设计药物的设计中应用 雌激素依赖性癌症的治疗。