A spectrophotometric procedure for rapid and sensitive measurements of beta-oxidation. Demonstration of factors that can be rate-limiting for beta-oxidation.

The Biochemical journal ·Vol. 164 ·No. 3 ·1977-06-15 ·Pages 621-33

Osmundsen H, Bremer J

Abstract

1. A spectrophotometric direct-reading assay for measurements of beta-oxidation by intact mitochondria is described. The procedure relies on the ability of ferricyanide to trap reducing equivalents generated by the acyl-CoA dehydrogenases (EC 1.3.99.3). The reduction of ferricyanide was recorded by using a dual-wavelength spectrophotometer. 2. Oxaloacetate or acetoacetate was used to stimulate the rate of beta-oxidation by rotenone-blocked mitochondria. Although both were effective with rat liver mitochondria, oxaloacetate gave about 75% more stimulation. With heart or kidney mitochondria, only oxaloacetate gave marked stimulation. Acetoacetate had no stimulatory effect with heart mitochondria, but a small stimulatory effect on the rate of beta-oxidation by kidney mitochondria. 3. The stoicheiometry of beta-oxidation-dependent reduction of ferricyanide was examined, and good correlations were found between experimental and theoretical amounts of ferricyanide reduced. 4. Ferricyanide appears as efficient a final electron acceptor as O2. With ferricyanide the rate of beta-oxidation by heart mitochondria can be measured without interference from the oxidation of tricarboxylic acid-cycle intermediates.

MeSH Terms

Acetoacetates/pharmacology Animals Carnitine/analogs & derivatives,pharmacology Ferricyanides Kinetics Male Mitochondria/drug effects,metabolism Oxaloacetates/pharmacology Oxidation-Reduction Oxidoreductases Proteins/metabolism Rats Rotenone/pharmacology Spectrophotometry/methods

Chemicals

Acetoacetates Ferricyanides Oxaloacetates Proteins Rotenone Oxidoreductases Carnitine

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Osmundsen H

Bremer J

References (22)

22 references, click to expand

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Article Info

Journal

The Biochemical journal

Abbr.

Biochem J

ISSN

0264-6021

DOI

10.1042/bj1640621

Published

1977-06-15

Pages

621-33

Language

English

Region

England

NLM ID

2984726R

PMCID

PMC1164840

Subset

External Links

PubMed source DOI: 10.1042/bj1640621 PMC Full Text View journal details

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A spectrophotometric procedure for rapid and sensitive measurements of beta-oxidation. Demonstration of factors that can be rate-limiting for beta-oxidation.

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