An acyl-coenzyme a dehydrogenase assay utilizing the ferricenium ion

Thomas C. Lehman, Daniel E. Hale, Ajay Bhala, Colin Thorpe

Research output: Contribution to journalArticlepeer-review

211 Scopus citations

Abstract

A sensitive assay for medium chain acyl-CoA dehydrogenase has been developed by substituting ferricenium hexafluorophosphate for the physiological acceptor, electron transferring flavoprotein. The ferricenium ion is a facile oxidant of the octanoyl-CoA-reduced enzyme with a Vmax of 1400 min-1 and a KM of 55 μm at pH 7.6. The ferricenium assay does not require additional mediator dyes, exhibits low background rates, and avoids the necessity of purifying substantial amounts of electron transferring flavoprotein. Unlike the fluorescence-based electron transferring flavoprotein assay, this new procedure can be performed aerobically. Both assays give comparable results when tested with crude fibroblast homogenates from normal and medium chain acyl-CoA dehydrogenase deficient patients. The convenience of the ferricenium method suggests it may be generally useful as a screening assay for a number of acyl-CoA dehydrogenases.

Original languageEnglish (US)
Pages (from-to)280-284
Number of pages5
JournalAnalytical Biochemistry
Volume186
Issue number2
DOIs
StatePublished - May 1 1990
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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