A Simple Spectrophotometric Assay for Long-Chain Acyl-CoA Dehydrogenase Activity Measurements in Human Skin Fibroblasts

Long-chain acyl-CoA dehydrogenase (LCAD) deficiency is an autosomal recessive disorder of fatty acid metabolism characterized by hypoglycemia, muscle weakness and hepato- and cardiomegaly to varying extents. Analysis of organic acids in urine usually reveals dicarboxylic aciduria with elevated levels of adipic, suberic and sebacic acids as well as longer chain dicarboxylic acids. Correct diagnosis of suspected patients requires measurement of LCAD in tissue or preferably, white blood cells and/or cultured skin fibroblasts. In this paper we present a simple spectrophotometric enzyme assay based on the use of ferricenium hexafluorophosphate as electron acceptor. Under optimized conditions the method presented allowed unequivocal identification of LCAD-deficiency in Fibroblast homogenates.

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Biochemical studies on cultured skin fibroblasts from a baby with long-chain acyl-CoA dehydrogenase deficiency presenting as sudden neonatal death

Journal of Inherited Metabolic Disease ◽

10.1007/bf01811420 ◽

1987 ◽

Vol 10 (S2) ◽

pp. 260-262 ◽

Cited By ~ 10

Author(s):

R. A. Chalmers ◽

N. English ◽

E. A. Hughes ◽

C. Noble-Jamieson ◽

J. S. Wigglesworth

Keyword(s):

Neonatal Death ◽

Dehydrogenase Deficiency ◽

Skin Fibroblasts ◽

Cultured Skin ◽

Biochemical Studies ◽

Chain Acyl ◽

Long Chain

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A new, simple assay for long-chain acyl-CoA dehydrogenase in cultured skin fibroblasts using stable isotopes and GC-MS

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease ◽

10.1016/0925-4439(92)90023-g ◽

1992 ◽

Vol 1180 (1) ◽

pp. 28-32 ◽

Cited By ~ 5

Author(s):

Klary E. Niezen-Koning ◽

Ronald J.A. Wanders ◽

Gils T. Nagel ◽

Lodewijk IJlst ◽

Hugo S.A. Heymans

Keyword(s):

Stable Isotopes ◽

Skin Fibroblasts ◽

Cultured Skin ◽

Chain Acyl ◽

Long Chain

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Adrenoleukodystrophy: Impaired Oxidation of Very Long Chain Fatty Acids in White Blood Cells, Cultured Skin Fibroblasts, and Amniocytes

Pediatric Research ◽

10.1203/00006450-198403000-00016 ◽

1984 ◽

Vol 18 (3) ◽

pp. 286-290 ◽

Cited By ~ 153

Author(s):

Inderjit Singh ◽

Ann E Moser ◽

Hugo W Moser ◽

Yasuo Kishimoto

Keyword(s):

Fatty Acids ◽

Blood Cells ◽

White Blood Cells ◽

Skin Fibroblasts ◽

Long Chain Fatty Acids ◽

Cultured Skin ◽

Long Chain ◽

Cells Cultured ◽

Chain Fatty Acids

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Very long-chain fatty acids in neutral lipids and glycerophospholipids of adrenoleukodystrophy-cultured skin fibroblasts

Journal of the Neurological Sciences ◽

10.1016/0022-510x(85)90068-1 ◽

1985 ◽

Vol 71 (2-3) ◽

pp. 301-306 ◽

Cited By ~ 10

Author(s):

Tetsuo Sakai ◽

Yasunobu Antoku ◽

Ikuo Goto ◽

Jun Ochiai ◽

Hiroshi Iwashita ◽

...

Keyword(s):

Fatty Acids ◽

Neutral Lipids ◽

Skin Fibroblasts ◽

Long Chain Fatty Acids ◽

Cultured Skin ◽

Long Chain ◽

Chain Fatty Acids

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Adrenoleukodystrophy: Impaired oxidation of long chain fatty acids in cultured skin fibroblasts and adrenal cortex

Biochemical and Biophysical Research Communications ◽

10.1016/s0006-291x(81)80142-8 ◽

1981 ◽

Vol 102 (4) ◽

pp. 1223-1229 ◽

Cited By ~ 100

Author(s):

Inderjit Singh ◽

Hugo W. Moser ◽

Ann B. Moser ◽

Yasuo Kishimoto

Keyword(s):

Fatty Acids ◽

Adrenal Cortex ◽

Skin Fibroblasts ◽

Long Chain Fatty Acids ◽

Cultured Skin ◽

Long Chain ◽

Chain Fatty Acids

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Improved determination of very-long-chain fatty acids in plasma and cultured skin fibroblasts: applications to the diagnosis of peroxisomal disorders

Journal of Chromatography B Biomedical Sciences and Applications ◽

10.1016/s0378-4347(00)82654-7 ◽

1989 ◽

Vol 494 ◽

pp. 31-41 ◽

Cited By ~ 21

Author(s):

W. Onkenhout ◽

P.F.H. van der Poel ◽

M.P.M. van den Heuvel

Keyword(s):

Fatty Acids ◽

Skin Fibroblasts ◽

Long Chain Fatty Acids ◽

Peroxisomal Disorders ◽

Cultured Skin ◽

Long Chain ◽

Chain Fatty Acids

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Determination of medium chain acyl-CoA dehydrogenase activity in cultured skin fibroblasts using mass spectrometry

Clinica Chimica Acta ◽

10.1016/0009-8981(91)90109-p ◽

1991 ◽

Vol 199 (2) ◽

pp. 173-184 ◽

Cited By ~ 9

Author(s):

K.E. Niezen-Koning ◽

T.E. Chapman ◽

I.E. Mulder ◽

G.P.A. Smit ◽

D.J. reijngoud ◽

...

Keyword(s):

Mass Spectrometry ◽

Dehydrogenase Activity ◽

Skin Fibroblasts ◽

Medium Chain ◽

Cultured Skin ◽

Chain Acyl

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Measurement of short-chain acyl-CoA dehydrogenase (SCAD) in cultured skin fibroblasts with hexanoyl-CoA as a competitive inhibitor to eliminate the contribution of medium-chain acyl-CoA dehydrogenase

Clinica Chimica Acta ◽

10.1016/0009-8981(94)90232-1 ◽

1994 ◽

Vol 229 (1-2) ◽

pp. 99-106 ◽

Cited By ~ 4

Author(s):

Klary E. Niezen-Koning ◽

Ronald J.A. Wanders ◽

Gijs T. Nagel ◽

Adrian C. Sewell ◽

Hugo S.A. Heymans

Keyword(s):

Competitive Inhibitor ◽

Skin Fibroblasts ◽

Short Chain ◽

Medium Chain ◽

Cultured Skin ◽

Chain Acyl

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Genetic Deficiency of Medium-Chain Acyl Coenzyme A Dehydrogenase: Studies in Cultured Skin Fibroblasts and Peripheral Mononuclear Leukocytes

Pediatric Research ◽

10.1203/00006450-198507000-00007 ◽

1985 ◽

Vol 19 (7) ◽

pp. 671-676 ◽

Cited By ~ 79

Author(s):

Paul M Coates ◽

Daniel E Hale ◽

Charles A Stanley ◽

Barbara E Corkey ◽

Jean A Cortner

Keyword(s):

Coenzyme A ◽

Skin Fibroblasts ◽

Mononuclear Leukocytes ◽

Medium Chain ◽

Cultured Skin ◽

Genetic Deficiency ◽

Chain Acyl ◽

Acyl Coenzyme A

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Metabolic Outcomes of Anaplerotic Dodecanedioic Acid Supplementation in Very Long Chain Acyl-CoA Dehydrogenase (VLCAD) Deficient Fibroblasts

Metabolites ◽

10.3390/metabo11080538 ◽

2021 ◽

Vol 11 (8) ◽

pp. 538

Author(s):

Igor Radzikh ◽

Erica Fatica ◽

Jillian Kodger ◽

Rohan Shah ◽

Ryan Pearce ◽

...

Keyword(s):

Alternative Energy ◽

Carbon Sources ◽

Krebs Cycle ◽

Dicarboxylic Acids ◽

High Energy ◽

Glycolytic Flux ◽

Chain Acyl ◽

Krebs Cycle Intermediates ◽

Dodecanedioic Acid ◽

Long Chain

Very long-chain acyl-CoA dehydrogenase deficiency (VLCADD, OMIM 609575) is associated with energy deficiency and mitochondrial dysfunction and may lead to rhabdomyolysis and cardiomyopathy. Under physiological conditions, there is a fine balance between the utilization of different carbon nutrients to maintain the Krebs cycle. The maintenance of steady pools of Krebs cycle intermediates is critical formitochondrial energy homeostasis especially in high-energy demanding organs such as muscle and heart. Even-chain dicarboxylic acids are established as alternative energy carbon sources that replenish the Krebs cycle by bypassing a defective β-oxidation pathway. Despite this, even-chain dicarboxylic acids are eliminated in the urine of VLCAD-affected individuals. In this study, we explore dodecanedioic acid (C12; DODA) supplementation and investigate its metabolic effect on Krebs cycle intermediates, glucose uptake, and acylcarnitine profiles in VLCAD-deficient fibroblasts. Our findings indicate that DODA supplementation replenishes the Krebs cycle by increasing the succinate pool, attenuates glycolytic flux, and reduces levels of toxic very long-chain acylcarnitines.

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