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

M. Kranendijk, G.S. Salomons, K.M. Gibson, C. Aktuglu-Zeybek, S. Bekri, E. Christensen, J. Clarke, A. Hahn, S.H. Korman, V. Mejaski-Bosnjak, A. Superti-Furga, C. Vianey-Saban, M.S. van der Knaap, C. Jakobs, E.A. Struys

Research output: Contribution to journalArticleAcademicpeer-review

8 Citations (Scopus)


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
Original languageEnglish
Pages (from-to)713-719
JournalJournal of Inherited Metabolic Disease
Issue number6
Publication statusPublished - 2009

Cite this