Abstract
Studies on the Mechanism of the Inhibition of Galactose Oxidation by Ethanol
Highlights
In an attempt to elucidate the mechanism of this inhibitory effect of ethanol on galactose metabolism by the liver, we have studied the influence of ethanol on the oxidation of galactosel-Cl4 to Cl402 in the soluble fractions of rat liver homogenates
Effect of Ethanol on Galactose-i-Cl4 Oxidation by Rat Liver Preparations-In experiments with the 105,000 x g supernatant fraction of rat liver homogenates, it was consistently observed that the addition of ethanol to the incubation system inhibited the osidation of galactose-1-C1402
The data obtained suggest that the inhibitory action of ethanol is caused by the DPNH generated in its oxidative metabolism
Summary
Materials and MethodsPreparation of Rat Liver Homogenatesand Incubation SystemFemale Sprague-Dawley rats (Charles River Breeding Laboratories, Inc., Boston, Massachusetts) weighing between 150 and 250 g were fasted overnight, killed by cervical dislocation, and the livers removed. Liver homogenates (1: 1, weight per volume) were prepared in ice-cold 0.15 M KC1 and 0.05 M nicotinamide with a Dounce homogenizer with a clearance of 0.5 to 1.0 mm. The soluble fraction was obtained by centrifugation at 105,000 x g in a Spinco preparative ultracentrifuge. The standard incubation medium used in the experiments consisted of 80 pmoles of potassium phosphate buffer, pH 7.4, 10 pmoles of ATP, 10 pmoles of MgC12, and 0.3 pmole of galactose-l-Cl4 (specific activity 0.8 pcurie per pmole). The protein in the soluble fraction of liver homogenates was measured by the biuret method [3]. The amount of protein added to each incubation was usually in the range of 50 mg. The total volume of the incubation medium, including all subsequent additions, was 1.6
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