A luminescence-based protocol for assessing fructose metabolism via quantification of ketohexokinase enzymatic activity in mouse or human hepatocytes.
STAR Protoc
; 2(3): 100731, 2021 09 17.
Article
em En
| MEDLINE
| ID: mdl-34409309
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).
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Ensaios Enzimáticos
/
Frutoquinases
/
Frutose
/
Medições Luminescentes
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article