A spectrophotometric procedure for rapid and sensitive measurements of β-oxidation. Demonstration of factors that can be rate-limiting for β-oxidation
- 15 June 1977
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 164 (3) , 621-633
- https://doi.org/10.1042/bj1640621
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.This publication has 23 references indexed in Scilit:
- Improved method for isolation of hypoglycins A and B from fruit of Blighia sapidaJournal of Pharmacy and Pharmacology, 1974
- On the mechanism of malonyl-CoA independent fatty acid synthesis. II. Isolation, properties and subcellular location of trans-2,3-hexenoyl-CoA and trans-2,3-decenoyl-CoA reductase.1972
- Localization of the Glycerol‐Phosphate Dehydrogenase in the Outer Phase of the Mitochondrial Inner MembraneEuropean Journal of Biochemistry, 1970
- The effect of pyrvate on cyclic oxidations by heart sarcosomesBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1967
- Effect of Antimycin A on Oxidative Phosphorylation with Ferricyanide as Electron Acceptor*Biochemistry, 1964
- Computer Programmes for Processing Enzyme Kinetic DataNature, 1963
- Preparation of crystalline lithium acetoacetateAnalytical Biochemistry, 1962
- The enzymic hydrolysis of adenosine triphosphate by liver mitochondria. 1. Activities at different pH valuesBiochemical Journal, 1957
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951
- Anaerobic oxidations. On ferricyanide as a reagent for the manometric investigation of dehydrogenase systemsBiochemical Journal, 1938