Faculty of Medicine, Institute of Biochemistry and Molecular Genetics, University of Ljubljana, Ljublijana, Slovenia.
Helmholtz Zentrum München, German Research Center for Environmental Health, Institute for Diabetes and Cancer, Neuherberg, Germany; German Center for Diabetes Research (DZD), Neuherberg, Germany.
Methods Enzymol. 2023;689:201-234. doi: 10.1016/bs.mie.2023.04.015. Epub 2023 May 20.
The 17β-hydroxysteroid dehydrogenase type 1 (HSD17B1) has a key role in estrogen biosynthesis as it catalyzes the reduction of estrone to the most potent estrogen, estradiol. Estradiol has a high affinity for estrogen receptors and thus stimulates their transactivation, which leads to cell proliferation and numerous other effects. HSD17B2 catalyzes the oxidation of estradiol to the less potent estrone, thereby decreasing estrogen receptor activation, which results in reduction of estrogen-associated effects. HSD17B1 and HSD17B2 overexpressing E.coli homogenates or recombinant enzymes can be used for screening and development of drugs against various pathologies such as cancer, endometriosis or osteoporosis. Here we describe the preparation of HSD17B1 and HSD17B2 bacterial homogenates and purified recombinant HSD17B1 protein as enzyme sources as well as enzymatic assays based on radiometric and mass-spectrometric detection for enzyme characterization.
17β-羟甾脱氢酶 1 型（HSD17B1）在雌激素生物合成中起着关键作用，因为它催化雌酮还原为最有效的雌激素，即雌二醇。雌二醇与雌激素受体具有高亲和力，从而刺激其转录激活，导致细胞增殖和许多其他效应。HSD17B2 催化雌二醇氧化为效力较低的雌酮，从而降低雌激素受体的激活，导致雌激素相关效应减少。HSD17B1 和 HSD17B2 过表达大肠杆菌匀浆或重组酶可用于筛选和开发针对各种疾病的药物，如癌症、子宫内膜异位症或骨质疏松症。在这里，我们描述了 HSD17B1 和 HSD17B2 细菌匀浆的制备以及作为酶源的纯化重组 HSD17B1 蛋白，以及基于放射性和质谱检测的酶特征的酶测定。
