A luminescence-based protocol for assessing fructose metabolism via quantification of ketohexokinase enzymatic activity in mouse or human hepatocytes.

Animals Carbohydrate Metabolism Enzyme Assays / methods Fructokinases / antagonists & inhibitors Fructose / metabolism Hepatocytes / enzymology Humans Luminescence Luminescent Measurements / methods Mice Non-alcoholic Fatty Liver Disease
Cell Biology Cell-based assays Metabolism

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
17 09 2021
Historique:
entrez: 19 8 2021
pubmed: 20 8 2021
medline: 20 8 2021
Statut: epublish

Résumé

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

Identifiants

DOI: 10.1016/j.xpro.2021.100731 PMID: 34409309 PMC: PMC8361265
pubmed: 34409309
doi: 10.1016/j.xpro.2021.100731
pii: S2666-1667(21)00438-X
pmc: PMC8361265
doi:

Substances chimiques

Fructose 30237-26-4
Fructokinases EC 2.7.1.-
ketohexokinase EC 2.7.1.3

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Pagination

100731

Informations de copyright

© 2021 The Author(s).

Déclaration de conflit d'intérêts

L.N. and H.-C.T. are employees of Alnylam Pharmaceuticals Inc. K.W., G.O.M., and J.B. are employees of AstraZeneca.

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Auteurs

Se-Hyung Park (SH)

Department of Pediatrics, Division of Pediatric Gastroenterology, University of Kentucky College of Medicine, 900 South Limestone, Wethington Rm 527, Lexington, KY 40536, USA.

Robert N Helsley (RN)

Department of Pediatrics, Division of Pediatric Gastroenterology, University of Kentucky College of Medicine, 900 South Limestone, Wethington Rm 527, Lexington, KY 40536, USA.

Leila Noetzli (L)

Alnylam Pharmaceuticals Inc., Cambridge, MA 021428, USA.

Ho-Chou Tu (HC)

Alnylam Pharmaceuticals Inc., Cambridge, MA 021428, USA.

Kristina Wallenius (K)

Bioscience, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden.

Gavin O'Mahony (G)

Medicinal Chemistry, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden.

Jeremie Boucher (J)

Bioscience, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden.
The Lundberg Laboratory for Diabetes Research, University of Gothenburg, Gothenburg, Sweden.
Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg, Gothenburg, Sweden.

Jianming Liu (J)

Discovery Biology, Discovery Sciences, R&D, AstraZeneca, Mölndal, Gothenburg 43183, Sweden.

Samir Softic (S)

Department of Pediatrics, Division of Pediatric Gastroenterology, University of Kentucky College of Medicine, 900 South Limestone, Wethington Rm 527, Lexington, KY 40536, USA.
Department of Pharmacology and Nutritional Sciences, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, MA, USA.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › A luminescence-based protocol for assessing...
questionsmedicales.fr › Métabolisme › Métabolisme glucidique › A luminescence-based protocol for assessing...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Dosages enzymatiques › A luminescence-based protocol for assessing...
questionsmedicales.fr › Enzymes et coenzymes › Enzymes › Transferases › Phosphotransferases › Phosphotransferases (Alcohol Group Acceptor) › Fructokinases › A luminescence-based protocol for assessing...
questionsmedicales.fr › Glucides › Sucres › Oses › Cétoses (sucres) › Fructose › A luminescence-based protocol for assessing...
questionsmedicales.fr › Cellules › Cellules épithéliales › Hépatocytes › A luminescence-based protocol for assessing...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › A luminescence-based protocol for assessing...
questionsmedicales.fr › Phénomènes physiques › Rayonnement › Rayonnement non ionisant › Lumière › Luminescence › A luminescence-based protocol for assessing...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Photométrie › Mesures de luminescence › A luminescence-based protocol for assessing...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › A luminescence-based protocol for assessing...
questionsmedicales.fr › Maladies de l'appareil digestif › Maladies du foie › Stéatose hépatique › Stéatose hépatique non alcoolique › A luminescence-based protocol for assessing...