Journal article

Publication year: 2009

Pages: 713-719

Journal: Journal of Inherited Metabolic Disease

Volume number: 32

Issue number: 6

ISSN: 0141-8955

eISSN: 1573-2665

DOI Link: https://doi.org/10.1007/s10545-009-1282-x

Publisher: Springer

Abstract: 
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.

Harvard Citation style: Kranendijk, M., Salomons, G., Gibson, K., Aktuglu-Zeybek, C., Bekri, S., Christensen, E., et al. (2009) 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, Journal of Inherited Metabolic Disease, 32(6), pp. 713-719. https://doi.org/10.1007/s10545-009-1282-x

APA Citation style: Kranendijk, M., Salomons, G., Gibson, K., Aktuglu-Zeybek, C., Bekri, S., Christensen, E., Clarke, J., Hahn, A., Korman, S., Mejaski-Bosnjak, V., Superti-Furga, A., Vianey-Saban, C., van der Knaap, M., Jakobs, C., & Struys, E. (2009). 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. Journal of Inherited Metabolic Disease. 32(6), 713-719. https://doi.org/10.1007/s10545-009-1282-x