Development and implementation of a novel assay for L-2-hydroxyglutarate dehydrogenase (L-2-HGDH) in cell lysates: L-2-HGDH deficiency in 15 patients with L-2-hydroxyglutaric aciduria.

Détails

ID Serval
serval:BIB_8851BD826773
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Development and implementation of a novel assay for L-2-hydroxyglutarate dehydrogenase (L-2-HGDH) in cell lysates: L-2-HGDH deficiency in 15 patients with L-2-hydroxyglutaric aciduria.
Périodique
Journal of Inherited Metabolic Disease
Auteur(s)
Kranendijk M., Salomons G.S., Gibson K.M., Aktuglu-Zeybek C., Bekri S., Christensen E., Clarke J., Hahn A., Korman S.H., Mejaski-Bosnjak V., Superti-Furga A., Vianey-Saban C., van der Knaap M.S., Jakobs C., Struys E.A.
ISSN
1573-2665 (Electronic)
ISSN-L
0141-8955
Statut éditorial
Publié
Date de publication
2009
Volume
32
Numéro
6
Pages
713-719
Langue
anglais
Résumé
L-2-hydroxyglutaric aciduria (L-2-HGA) is a rare inherited autosomal recessive neurometabolic disorder caused by mutations in the gene encoding L-2-hydroxyglutarate dehydrogenase. An assay to evaluate L-2-hydroxyglutarate dehydrogenase (L-2-HGDH) activity in fibroblast, lymphoblast and/or lymphocyte lysates has hitherto been unavailable. We developed an L-2-HGDH enzyme assay in cell lysates based on the conversion of stable-isotope-labelled L-2-hydroxyglutarate to 2-ketoglutarate, which is converted into L-glutamate in situ. The formation of stable isotope labelled L-glutamate is therefore a direct measure of L-2-HGDH activity, and this product is detected by liquid chromatography-tandem mass spectrometry. A deficiency of L-2-HGDH activity was detected in cell lysates from 15 out of 15 L-2-HGA patients. Therefore, this specific assay confirmed the diagnosis unambiguously affirming the relationship between molecular and biochemical observations. Residual activity was detected in cells derived from one L-2-HGA patient. The L-2-HGDH assay will be valuable for examining in vitro riboflavin/FAD therapy to rescue L-2-HGDH activity.
Mots-clé
Alcohol Oxidoreductases/analysis, Alcohol Oxidoreductases/cerebrospinal fluid, Animals, Brain Diseases, Metabolic, Inborn/cerebrospinal fluid, Brain Diseases, Metabolic, Inborn/diagnosis, Calibration, Cell Extracts/analysis, Cell Extracts/chemistry, Cells, Cultured, Chromatography, High Pressure Liquid, Chromatography, Liquid/methods, Enzyme Assays/methods, Enzyme Assays/standards, Fibroblasts/chemistry, Fibroblasts/enzymology, Humans, Lymphocytes/chemistry, Lymphocytes/enzymology, Models, Biological, Models, Molecular, Rats, Research Design, Tandem Mass Spectrometry/methods
Pubmed
Web of science
Création de la notice
14/03/2011 17:08
Dernière modification de la notice
20/08/2019 15:47
Données d'usage