Evaluation of electron-transfer flavoprotein and alpha-lipoamide dehydrogenase redox states by two-channel fluorimetry and its application to the investigation of beta-oxidation.
A new method permitting the simultaneous evaluation of the redox states of alpha-lipoamide dehydrogenase and electron-transfer flavoprotein in intact rat liver mitochondria by two-channel fluorimetry is described. It is shown that correction for the partial overlap of emission spectra can readily be introduced after a calibration procedure is performed. This method was applied to the investigation into regulation of palmitoylcarnitine oxidation. It was found that in the presence of rotenone, malonate and a redox buffer for the mitochondrial NAD-system, the beta-oxidation flux was sensitive to variations in redox state of respiratory chain electron carriers at low states of NAD reduction. Therefore, the concept of beta-oxidation control caused solely by the NAD redox state can no longer be sustained.
Kunz
W S
WS
Institut für Biochemie, Medizinischen Akademie Magdeburg, G.D.R.
D000818:Animals; D008058:Dihydrolipoamide Dehydrogenase / Q000378:metabolism*; D004579:Electron Transport; D005420:Flavoproteins / Q000378:metabolism*; D007700:Kinetics; D008930:Mitochondria, Liver / Q000201:enzymology*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D051381:Rats; D013050:Spectrometry, Fluorescence / Q000379:methods
D005420:Flavoproteins; D008058:Dihydrolipoamide Dehydrogenase