A new, simple assay for long-chain acyl-CoA dehydrogenase in cultured skin fibroblasts using stable isotopes and GC-MS.
In this paper, we present a new method for measurement of long-chain acyl-CoA dehydrogenase (LCAD) activities in cultured skin fibroblasts. The method is based upon gas chromatographic/mass spectrometric determination of 3-OH-hexadecanoic acid formed during incubation of fibroblasts in a medium containing palmitoyl-CoA and crotonase, to convert the enoyl-CoA ester produced into the 3-hydroxyacyl-CoA ester. The validity of the method is demonstrated by the finding of a full deficiency of LCAD in fibroblasts from three patients with an established deficiency of LCAD.
Niezen-Koning
K E
KE
Department of Pediatrics, University Hospital Groningen, The Netherlands.
Wanders
R J
RJ
Nagel
G T
GT
IJlst
L
L
Heymans
H S
HS
D042964:Acyl-CoA Dehydrogenase; D044942:Acyl-CoA Dehydrogenase, Long-Chain / Q000032:analysis* / Q000172:deficiency; D002478:Cells, Cultured; D004746:Enoyl-CoA Hydratase / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D007554:Isotopes; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000201:enzymology*
D007554:Isotopes; D010171:Palmitoyl Coenzyme A; D042964:Acyl-CoA Dehydrogenase; D044942:Acyl-CoA Dehydrogenase, Long-Chain; D004746:Enoyl-CoA Hydratase