title: A new, simple assay for long-chain acyl-CoA dehydrogenase in cultured skin fibroblasts using stable isotopes and GC-MS.
abstract: 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.
authors.affiliation: Department of Pediatrics, University Hospital Groningen, The Netherlands.
authors.forename: K E
authors.initials: KE
authors.lastname: Niezen-Koning
authors.forename: R J
authors.initials: RJ
authors.lastname: Wanders
authors.forename: G T
authors.initials: GT
authors.lastname: Nagel
authors.forename: L
authors.initials: L
authors.lastname: IJlst
authors.forename: H S
authors.initials: HS
authors.lastname: Heymans
mesh_terms: 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*
chemical_list: D007554:Isotopes; D010171:Palmitoyl Coenzyme A; D042964:Acyl-CoA Dehydrogenase; D044942:Acyl-CoA Dehydrogenase, Long-Chain; D004746:Enoyl-CoA Hydratase