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; Ernst 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

doi:10.1007/s10545-009-1282-x

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, Ernst 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

Institut for Klinisk Medicin

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

9 Citationer (Scopus)

Abstract

Udgivelsesdato: 2009-Dec

Originalsprog	Engelsk
Tidsskrift	Journal of Inherited Metabolic Disease
Vol/bind	32
Udgave nummer	6
Sider (fra-til)	713-9
Antal sider	6
ISSN	0141-8955
DOI	https://doi.org/10.1007/s10545-009-1282-x
Status	Udgivet - 2009

Bibliografisk note

Keywords: Alcohol Oxidoreductases; Animals; Brain Diseases, Metabolic, Inborn; Calibration; Cell Extracts; Cells, Cultured; Chromatography, High Pressure Liquid; Chromatography, Liquid; Enzyme Assays; Fibroblasts; Humans; Lymphocytes; Models, Biological; Models, Molecular; Rats; Research Design; Tandem Mass Spectrometry

Adgang til dokumentet

10.1007/s10545-009-1282-x

Citationsformater

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. (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), 713-9. https://doi.org/10.1007/s10545-009-1282-x

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. / Kranendijk, M; Salomons, G S; Gibson, K M et al.
I: Journal of Inherited Metabolic Disease, Bind 32, Nr. 6, 2009, s. 713-9.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

Kranendijk, M, Salomons, GS, Gibson, KM, Aktuglu-Zeybek, C, Bekri, S, Christensen, E, Clarke, J, Hahn, A, Korman, SH, Mejaski-Bosnjak, V, Superti-Furga, A, Vianey-Saban, C, van der Knaap, MS, Jakobs, C & Struys, EA 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, bind 32, nr. 6, s. 713-9. https://doi.org/10.1007/s10545-009-1282-x

@article{21e911c072d411df928f000ea68e967b,

title = "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",

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.",

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

note = "Keywords: Alcohol Oxidoreductases; Animals; Brain Diseases, Metabolic, Inborn; Calibration; Cell Extracts; Cells, Cultured; Chromatography, High Pressure Liquid; Chromatography, Liquid; Enzyme Assays; Fibroblasts; Humans; Lymphocytes; Models, Biological; Models, Molecular; Rats; Research Design; Tandem Mass Spectrometry",

year = "2009",

doi = "10.1007/s10545-009-1282-x",

language = "English",

volume = "32",

pages = "713--9",

journal = "Journal of Inherited Metabolic Disease",

issn = "0141-8955",

publisher = "Springer",

number = "6",

}

TY - JOUR

T1 - 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

AU - Kranendijk, M

AU - Salomons, G S

AU - Gibson, K M

AU - Aktuglu-Zeybek, C

AU - Bekri, S

AU - Christensen, Ernst

AU - Clarke, J

AU - Hahn, A

AU - Korman, S H

AU - Mejaski-Bosnjak, V

AU - Superti-Furga, A

AU - Vianey-Saban, C

AU - van der Knaap, M S

AU - Jakobs, C

AU - Struys, E A

N1 - Keywords: Alcohol Oxidoreductases; Animals; Brain Diseases, Metabolic, Inborn; Calibration; Cell Extracts; Cells, Cultured; Chromatography, High Pressure Liquid; Chromatography, Liquid; Enzyme Assays; Fibroblasts; Humans; Lymphocytes; Models, Biological; Models, Molecular; Rats; Research Design; Tandem Mass Spectrometry

PY - 2009

Y1 - 2009

N2 - 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.

AB - 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.

U2 - 10.1007/s10545-009-1282-x

DO - 10.1007/s10545-009-1282-x

M3 - Journal article

C2 - 19821142

SN - 0141-8955

VL - 32

SP - 713

EP - 719

JO - Journal of Inherited Metabolic Disease

JF - Journal of Inherited Metabolic Disease

IS - 6

ER -