Purification and characterization of human testis aldose and aldehyde reductase.
1. Aldose reductase and aldehyde reductase were purified to homogeneity from human testis. 2. The molecular weight of aldose reductase and aldehyde reductase were estimated to be 36,000 and 38,000 by SDS-PAGE, and the pI values of these enzymes were found to be 5.9 and 5.1 by chromatofocusing, respectively. 3. Aldose reductase had activity for aldo-sugars, whereas aldehyde reductase was virtually inactive for aldo-sugars. The Km values of aldose reductase for D-glucose, D-galactose and D-xylose were 57, 49 and 6.2 mM, respectively. Aldose reductase utilized both NADPH and NADH as coenzymes, whereas aldehyde reductase only NADPH. 4. Sulfate ion caused 3-fold activation of aldose reductase, but little for that of aldehyde reductase. 5. Sodium valproate inhibited significantly aldehyde reductase, but not aldose reductase. Aldose reductase was inhibited strongly by aldose reductase inhibitors being in clinical trials at concentrations of the order of 10(-7)-10(-9) M. Aldehyde reductase was also inhibited by these inhibitors, but its susceptibility was less than aldose reductase. 6. Reaction of aldose reductase with pyridoxal 5'-phosphate (PLP) resulted ca 2.5-fold activation, but aldehyde reductase did not cause the activation. PLP-treated aldose reductase has lost the susceptibility to aldose reductase inhibitor.
Tanimoto
T
T
Division of Biological Chemistry, National Institute of Hygienic Sciences, Tokyo, Japan.
Ohta
M
M
Tanaka
A
A
Ikemoto
I
I
Machida
T
T
D000426:Alcohol Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D007526:Isoelectric Point; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D013379:Substrate Specificity; D013737:Testis / Q000201:enzymology*
D011732:Pyridoxal Phosphate; D000426:Alcohol Dehydrogenase; D000449:Aldehyde Reductase