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

Biochim Biophys Acta. 1992 Oct 13;1180(1):28-32. doi: 10.1016/0925-4439(92)90023-g.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyl-CoA Dehydrogenase
  • Acyl-CoA Dehydrogenase, Long-Chain / analysis*
  • Acyl-CoA Dehydrogenase, Long-Chain / deficiency
  • Cells, Cultured
  • Enoyl-CoA Hydratase / metabolism
  • Fibroblasts / enzymology
  • Gas Chromatography-Mass Spectrometry / methods*
  • Humans
  • Isotopes
  • Palmitoyl Coenzyme A / metabolism
  • Skin / cytology
  • Skin / enzymology*

Substances

  • Isotopes
  • Palmitoyl Coenzyme A
  • Acyl-CoA Dehydrogenase
  • Acyl-CoA Dehydrogenase, Long-Chain
  • Enoyl-CoA Hydratase