Abstract
Development of the use of flavin and nicotinamide-adenine nucleotide fluorescence in monitoring the redox state of the free mitochondrial NADH/NAD+couple in cells, tissues and organs is reviewed. A break-through was the identification of dihydrolipoamide dehydrogenase (FpL) as the major NAD-linked fluorescent flavoprotein of mitochondria. This mitochondrial matrix flavoprotein is in equilibrium with the free NADH/NAD+pool and its mid-potential is sufficiently near to that of NADH/NAD+so that its percentage reduction follows that of the latter. Possibilities of monitoring mitochondrial and cytosolic NADH depend on the population density of mitochondria and thus are tissue-dependent. Upon a shift toward reduction, fluorescence intensities of NADH and flavins swing to reciprocal directions, so that the NADH/flavin fluorescence ratio can be used to increase the sensitivity of redox monitoring. This method is attaining widening use in studies on metabolic regulation under normal and pathological conditions.
Highlights
Development of the use of °avin and nicotinamide-adenine nucleotide °uorescence in monitoring the redox state of the free mitochondrial NADH/NADþ couple in cells, tissues and organs is reviewed
This mitochondrial matrix °avoprotein is in equilibrium with the free NADH/NADþ pool and its mid-potential is su±ciently near to that of NADH/NADþ so that its percentage reduction follows that of the latter
Further studies on rat liver mitochondria and submitochondrial particles at the Johnson Foundation, revealed, that the high-°uorescence, low-potential °avin (FpF1) resides in the soluble mitochondrial matrix, whereas the low-°uorescence, NADH-linked °avin with higher redox potential was membrane bound
Summary
From identication of °uorescent °avoproteins to mitochondrial redox indicators in intact tissues Development of the use of °avin and nicotinamide-adenine nucleotide °uorescence in monitoring the redox state of the free mitochondrial NADH/NADþ couple in cells, tissues and organs is reviewed.
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