Michaelis-Menten Kinetics Measurements of Aldo-Keto Reductases for Various Substrates in Murine Tissue

STAR Protoc. 2020 Dec 7;1(3):100206. doi: 10.1016/j.xpro.2020.100206. eCollection 2020 Dec 18.

Abstract

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).

Keywords: Metabolism; Molecular Biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Reductase / chemistry
  • Aldehyde Reductase / genetics
  • Aldo-Keto Reductases / genetics
  • Aldo-Keto Reductases / metabolism
  • Aldo-Keto Reductases / pharmacokinetics*
  • Animals
  • Enzyme Assays / methods*
  • Enzymes / metabolism*
  • Kinetics
  • Malondialdehyde
  • Mice
  • Pyruvaldehyde / metabolism
  • Substrate Specificity

Substances

  • Enzymes
  • Malondialdehyde
  • Pyruvaldehyde
  • Aldo-Keto Reductases
  • Aldehyde Reductase