Rat aldose reductase-like protein (AKR1B14) efficiently reduces the lipid peroxidation product 4-oxo-2-nonenal.
In this study, we examined the substrate specificity, inhibitor sensitivity and kinetic mechanism of a rat aldose reductase-like protein, which is named AKR1B14 in the aldo-keto reductase (AKR) superfamily. AKR1B14 catalyzed the nicotinamide adenine dinucleotide phosphate reduced form (NADPH)-dependent reduction of carbonyl compounds (derived from lipid peroxidation and glycation), xenobiotic aromatic aldehydes and some aromatic ketones. 4-Oxo-2-nonenal, the best substrate showing a K(m) value of 0.16 µM, was reduced into less reactive 4-oxo-2-nonenol, and its cytotoxicity was attenuated by the overexpression of the enzyme in cultured cells. The enzyme also showed low K(m) values (0.9-10 µM) for medium-chain aliphatic aldehydes (such as 4-hydroxynonenal, 1-hexenal and farnesal) and 3-deoxyglucosone, although the K(m) values for short-chain substrates (such as isocaproaldehyde, acrolein and methylglyoxal) were high (16-600 µM). In the reverse reaction, aliphatic and aromatic alcohols were oxidized by AKR1B14 at low rates. AKR1B14 was inhibited by aldose reductase inhibitors such as tolrestat and epalrestat, and their inhibition patterns were noncompetitive versus the aldehyde substrate and competitive with respect to the alcohol substrate. Kinetic analyses of the oxidoreduction and dead-end inhibition suggest that the reaction follows an ordered sequential mechanism.
Laboratory of Biochemistry, Gifu Pharmaceutical University, Daigaku-Nishi, Gifu 501–1196, Japan. sendo@gifu-pu.ac.jp
Satoshi
S
Endo
Toshiyuki
T
Matsunaga
Anna
A
Fujita
Kazuo
K
Tajima
Ossama
O
El-Kabbani
Akira
A
Hara
D000429:Alcohol Oxidoreductases / Q000378:metabolism; D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism*; D000074408:Aldo-Keto Reductases; D000818:Animals; D055162:Biocatalysis; D002417:Cattle; D006801:Humans; D015227:Lipid Peroxidation; D008099:Liver / Q000201:enzymology*; D009249:NADP / Q000378:metabolism; D018384:Oxidative Stress; D051381:Rats; D013379:Substrate Specificity
C403894:4-oxo-2-nonenal; D000447:Aldehydes; D009249:NADP; D000429:Alcohol Oxidoreductases; C558977:Akr1b7 protein, rat; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase