Department of Internal Medicine I and Clinical Chemistry, University Hospital Heidelberg, Heidelberg 69120, Germany.
German Center for Diabetes Research (DZD), Neuherberg 85764, Germany.
STAR Protoc. 2020 Dec 7;1(3):100206. doi: 10.1016/j.xpro.2020.100206. eCollection 2020 Dec 18.
Aldo-keto reductases (AKRs) are responsible for the detoxification of harmful aldehydes. Due to the large number of isotypes, the physiological relevance of AKRs cannot be obtained using mRNA or protein quantification, but only through the use of enzymatic assays to demonstrate functionality. Here, we present a fast and simple protocol to determine the important Michaelis-Menten kinetics of AKRs, which includes various aldehyde substrates of interest such as 4-hydroxynonenal, methylglyoxal, and malondialdehyde. For complete details on the use and execution of this protocol, please refer to Morgenstern et al. (2017) and Schumacher et al. (2018).
醛酮还原酶（AKRs）负责解毒有害的醛类。由于同工酶数量众多，因此无法通过 mRNA 或蛋白质定量来获得 AKR 的生理相关性，而只能通过酶促测定来证明其功能。在这里，我们提出了一种快速而简单的方案来确定 AKR 的重要米氏动力学，其中包括各种感兴趣的醛类底物，如 4-羟基壬烯醛、甲基乙二醛和丙二醛。有关此方案的使用和执行的完整详细信息，请参见 Morgenstern 等人（2017 年）和 Schumacher 等人（2018 年）。
