Characterization of aldose reductase and aldehyde reductase from rat testis.
Aldose reductase (alditol:NAD(P)+ 1-oxidoreductase, EC 1.1.1.21) and aldehyde reductase (alcohol:NADP+ oxidoreductase, EC 1.1.1.2) were purified to a homogeneity from rat testis. The molecular weights of aldose reductase and aldehyde reductase were estimated to be 38,000 and 41,000 by SDS-polyacrylamide gel electrophoresis, and the pI values of these enzymes were found to be 5.3 and 6.1 by chromatofocusing, respectively. Aldose reductase had activity for aldo-sugars such as xylose, glucose and galactose, whereas aldehyde reductase was virtually inactive for these aldo-sugars. The Km values of aldose reductase for aldo-sugars were relatively high. When a correction was made for the fraction of aldo-sugar present as the aldehyde form, which is the real substrate of the enzyme, the Km values were much lower. Aldose reductase utilized both NADPH and NADH as coenzyme, whereas aldehyde reductase utilized only NADPH. Aldose reductase was activated significantly by sulfate ion, while aldehyde reductase was little affected. Both enzymes were inhibited strongly by the known aldose reductase inhibitors. However, aldehyde reductase was in general less susceptible to these inhibitors when compared to aldose reductase. Both aldose reductase and aldehyde reductase treated with pyridoxal 5-phosphate have lost the susceptibility to aldose reductase inhibitor, suggesting that in these two enzymes aldose reductase inhibitor interacts with a lysine residue.
Kawasaki
N
N
Division of Biological Chemistry, National Institute of Hygienic Sciences, Tokyo, Japan.
Tanimoto
T
T
Tanaka
A
A
D000426:Alcohol Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D007526:Isoelectric Point; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D010658:Phenylglyoxal / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D051381:Rats; D012492:Salts / Q000494:pharmacology; D013379:Substrate Specificity; D013401:Sugar Alcohol Dehydrogenases / Q000378:metabolism*; D013737:Testis / Q000201:enzymology*
D012492:Salts; D011732:Pyridoxal Phosphate; D013401:Sugar Alcohol Dehydrogenases; D000426:Alcohol Dehydrogenase; D000449:Aldehyde Reductase; D004047:Diethyl Pyrocarbonate; D010658:Phenylglyoxal