Naslov
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

Autori
Kranendijk, M. ; Salomons, G.S. ; Gibson, K.M. ; Aktuglu-Zeybek, C. ; Bekri, S. ; Christensen, E. ; Clarke, J. ; Hahn, A. ; Korman, S.H. ; Mejaški-Bošnjak, Vlatka ; Superti-Furga, A. ; Vianey-Saban, C. ; van der Knaap, M.S. ; Jakobs, C. ; Struys, E.A.

Izvornik
Journal of inherited metabolic disease (0141-8955) 32 (2009), 6; 713-719

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
L-2-hydroxyglutarate dehydrogenase; L-2-hydroxyglutaric aciduria; Assay

Sažetak
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.

Izvorni jezik
Engleski

Znanstvena područja
Kliničke medicinske znanosti

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