title: A functional arginine residue in NADPH-dependent aldehyde reductase from pig kidney.
abstract: Pig kidney aldehyde reductase is inactivated by 2,3-butanedione, phenylglyoxal, methylglyoxal, and 1,2-cyclohexanedione. 2,3-Butanedione caused the most rapid loss in enzyme activity, the rate of loss being proportional to the concentration of 2,3-butanedione. Neither D-glyceraldehyde nor pyridine 3-aldehyde, both substrates for this broadly specific enzyme, protected the enzyme from inactivation but 1 mM NADPH or NADP completely prevented the loss of activity by 2,3-butanedione suggesting the involvement of arginine in the binding of cofactor. Nicotinamide mononucleotide (NMN) (reduced form) offered no protection to inactivation whereas ADP-ribose phosphate gave complete protection indicating that it is the latter portion of NADPH which interacts with the essential arginine. Both NMN and ADP-ribose phosphate are competitive inhibitors of aldehyde reductase with respect to NADPH. Butanedione-modified aldehyde reductase could still bind to a blue dextran-Sepharose 4B column suggesting that the modified arginine did not bind NADPH. This was confirmed by fluorescence spectra which showed that chemically modified aldehyde reductase caused the same blue shift of NADPH fluorescence as did native aldehyde reductase. Of additional interest was the quenching of NADPH fluorescence by aldehyde reductase which, with one exception, is in contrast to the fluorescence behavior of all other oxidoreductases.
mesh_terms: D000445:Aldehyde Oxidoreductases / Q000378:metabolism*; D000818:Animals; D001120:Arginine; D003931:Diacetyl / Q000494:pharmacology; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D009249:NADP; D009537:Nicotinamide Mononucleotide / Q000494:pharmacology; D011485:Protein Binding; D011765:Pyruvaldehyde / Q000494:pharmacology; D013053:Spectrophotometry; D013552:Swine
chemical_list: D009537:Nicotinamide Mononucleotide; D009249:NADP; D011765:Pyruvaldehyde; D001120:Arginine; D000445:Aldehyde Oxidoreductases; D003931:Diacetyl