An acyl-coenzyme a dehydrogenase assay utilizing the ferricenium ion

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

Research output: Contribution to journalArticle

207 Citations (Scopus)

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

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Electron-Transferring Flavoproteins
Coenzymes
Acyl-CoA Dehydrogenase
Assays
Oxidoreductases
Ions
Acyl-CoA Dehydrogenases
Oxidants
Coloring Agents
Fibroblasts
Fluorescence
Enzymes
Screening

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lehman, Thomas C. ; Hale, Daniel ; Bhala, Ajay ; Thorpe, Colin. / An acyl-coenzyme a dehydrogenase assay utilizing the ferricenium ion. In: Analytical Biochemistry. 1990 ; Vol. 186, No. 2. pp. 280-284.
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An acyl-coenzyme a dehydrogenase assay utilizing the ferricenium ion. / Lehman, Thomas C.; Hale, Daniel; Bhala, Ajay; Thorpe, Colin.

In: Analytical Biochemistry, Vol. 186, No. 2, 01.05.1990, p. 280-284.

Research output: Contribution to journalArticle

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