Covalent binding of acetaldehyde selectively inhibits the catalytic activity of lysine-dependent enzymes.
Hepatic ethanol metabolism generates the reactive intermediate, acetaldehyde, which binds to proteins. The binding of acetaldehyde to purified enzymes was determined in order to ascertain whether such binding altered their catalytic functions. [14C]Acetaldehyde was incubated with alcohol dehydrogenase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase and RNase A, each at 37 degrees C (pH 7.4). In some reactions, sodium cyanoborohydride was included for stabilization of Schiff bases, formed as a result of the reaction between acetaldehyde and the amino groups of the enzymes. Portions of each reaction mixture were removed for determination of stable and total (stable plus borohydride-reducible) adducts. Alcohol dehydrogenase and lactate dehydrogenase were not inhibited by adduct formation. Glucose-6-phosphate dehydrogenase and RNase, the activities of which depend on a lysine residue at their catalytic sites, were inhibited in a dose- and time-dependent manner. The degree of inhibition directly correlated with total adduct formation. Phosphate, known to inhibit binding to the active site lysine of RNase, prevented the inhibition of catalytic activity caused by adduct formation. These findings indicate that the binding of acetaldehyde to lysine at the catalytic site can inhibit enzyme activity.
D000079:Acetaldehyde / Q000378:metabolism*; D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D011485:Protein Binding; D012259:Ribonuclease, Pancreatic / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; D007770:L-Lactate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D012259:Ribonuclease, Pancreatic; D000079:Acetaldehyde; D008239:Lysine