Polyol-pathway enzymes of human brain. Partial purification and properties of aldose reductase and hexonate dehydrogenase.
Aldose reductase and hexonate dehydrogenase were isolated from human brain and partially purified. The two enzymes exhibited distinctive substrate-specificity profiles with a variety of aldoses,and aliphatic and aromatic aldehydes. Aldose reductase exhibited a high affinity for DL-glyceraldehyde (Km of 62 microM) and a low affinity (Km of 90 mM) for glucose, the physiological substrate of the polyol pathway. Hexonate dehydrogenase exhibited a relatively low affinity for D-glucuronate (Km of 4.6 mM) and a very low affinity for glucose (Km of 390 mM). Both enzymes exhibited a high specificity for NADPH, and both were inhibited competitively by NADP+. Hexonate dehydrogenase was inhibited by iodoacetate, iodoacetamide, N-ethylmaleimide and p-chloromercuribenzoate. Preincubation with 2-mercaptoethanol resulted in activation. Both enzymes were inhibited by a number of barbiturates (barbital, phenobarbital and pentobarbital) and by the central-nervous-system drugs diphenylhydantoin and ethosuccinimide. The substrate specificity and pattern of inhibition suggest that the two enzymes isolated correspond to two of four previously reported aldehyde reductases isolated from human brain.
D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D001921:Brain / Q000201:enzymology*; D002237:Carbohydrate Dehydrogenases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D002848:Chromatography, DEAE-Cellulose; D005965:Glucuronates / Q000037:antagonists & inhibitors / Q000302:isolation & purification; D005990:Glycerol / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D009249:NADP / Q000378:metabolism; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000302:isolation & purification*; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D005965:Glucuronates; D013439:Sulfhydryl Reagents; D009249:NADP; D002237:Carbohydrate Dehydrogenases; D013401:Sugar Alcohol Dehydrogenases; C020590:L-glucuronate reductase; D000449:Aldehyde Reductase; D005990:Glycerol