Detlefsen Andrea J
Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States.
Methods Enzymol. 2023;689:303-329. doi: 10.1016/bs.mie.2023.04.007. Epub 2023 Apr 25.
Aldo-keto reductase family 1C (AKR1C) members transform steroids via their 3-, 17-, and 20-ketosteroid reductase activities. The biochemical study of these enzymes can help to inform their roles in hormone-dependent diseases and develop therapeutic inhibitors. This work describes a protocol to purify AKR1C1-4 members from a bacterial expression system using two chromatography steps. It also describes the basis of discontinuous assays to measure steroid conversion.
醛糖酮还原酶家族1C（AKR1C）成员通过其3-、17-和20-酮类固醇还原酶活性转化类固醇。对这些酶的生化研究有助于了解它们在激素依赖性疾病中的作用，并开发治疗性抑制剂。这项工作描述了一种使用两步色谱法从细菌表达系统中纯化AKR1C1-4成员的方案。它还描述了用于测量类固醇转化的不连续测定法的基础。
