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Updated: Oct 23, 2025
Determination of Fatty Acid Oxidation and Lipogenesis in Mouse Primary Hepatocytes
Published on: August 27, 2015
A luminescence-based protocol for assessing fructose metabolism via quantification of ketohexokinase enzymatic
Se-Hyung Park1, Robert N Helsley1, Leila Noetzli2
1Department of Pediatrics, Division of Pediatric Gastroenterology, University of Kentucky College of Medicine, 900 South Limestone, Wethington Rm 527, Lexington, KY 40536, USA.
Abstract:
Ketohexokinase (KHK) catalyzes the first step of fructose metabolism. Inhibitors of KHK enzymatic activity are being evaluated in clinical trials for the treatment of non-alcoholic fatty liver disease (NAFLD) and diabetes. Here, we present a luminescence-based protocol to quantify KHK activity. The accuracy of this technique has been validated using knockdown and overexpression of KHK in vivo and in vitro. The specificity of the assay has been verified using 3-O-methyl-D-fructose, a non-metabolizable analog of fructose, heat inactivation of hexokinases, and depletion of potassium. For complete details on the use of this protocol, please refer to Damen et al. (2021).
