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).
|State||Published - Sep 17 2021|
Bibliographical note© 2021 The Author(s).
- Carbohydrate Metabolism
- Enzyme Assays/methods
- Fructokinases/antagonists & inhibitors
- Luminescent Measurements/methods
- Non-alcoholic Fatty Liver Disease