The binding of dihydronicotinamide--adenine dinucleotide and pyridine-3-aldehyde--adenine dinucleotide by yeast alcohol dehydrogenase.
1. Yeast alcohol dehydrogenase has been found to react with NADH in the presence of acetamide to form a highly fluorescent ternary complex. Titration of the enzyme to form this complex has provided a method for the estimation of the number of binding sites on the enzyme. 2. The binding of NADH by the enzyme has been studied independently, with a modified form of equilibrium dialysis, by using gel filtration. 3. A third method, depending upon the formation of a ternary complex of enzyme, hydroxylamine and pyridine-3-aldehyde-adenine dinucleotide, has also been used to titrate the enzyme. 4. Values obtained with all three methods are substantially in agreement that only three coenzyme-binding sites are available. This is in contrast with the established fact that the enzyme is composed of four identical subunits.
D000227:Adenine Nucleotides / Q000378:metabolism*; D000429:Alcohol Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism*; D000577:Amides / Q000378:metabolism; D001665:Binding Sites; D003956:Dialysis
D000227:Adenine Nucleotides; D000447:Aldehydes; D000577:Amides; D000429:Alcohol Oxidoreductases