Nitric oxide-independent, thiol-associated ADP-ribosylation inactivates aldehyde dehydrogenase.
Nitric oxide inhibits the activity of glyceraldehyde-3-phosphate dehydrogenase and stimulates NAD-dependent automodification of a cysteine (Dimmeler, S., Lottspeich, F., and Brüne, B. (1992) J. Biol. Chem. 267, 16771-16774). Another NAD-utilizing dehydrogenase that has a catalytic cysteine, aldehyde dehydrogenase (ALDH), was also inhibited by nitric oxide. Unlike glyceraldehyde-3-phosphate dehydrogenase, ALDH was modified in a nitric oxide-independent process by ADP-ribose, but not by NAD. Modification, which proceeded to > 2 mol ADP-ribose.mol ALDH-1, was associated with an exponential decrease in enzyme activity to less than 10% of control. Two types of evidence suggested modification of the ALDH-active site: 1) ADP-ribose inhibited ALDH competitively (Ki = 0.46 mM) with respect to NAD (Km = 0.11 mM) in brief incubations and 2) the presence of substrates protected ALDH from both modification and inhibition by ADP-ribose. The ALDH-ADP-ribose bond was sensitive to base and mercuric ion and stable to acid and neutral hydroxylamine, properties shared with the ADP-ribosylcysteine linkage synthesized enzymatically by pertussis toxin. These data demonstrate a novel means of inactivation of an NAD-dependent enzyme, namely the affinity-based modification of the enzyme NAD site by ADP-ribose, and suggest that nonenzymatic ADP-ribosylation may be responsible for modification of cysteine residues.
McDonald
L J
LJ
Laboratory of Cellular Metabolism, National Institutes of Health, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892.
Moss
J
J
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001345:Autoradiography; D001921:Brain / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000037:antagonists & inhibitors / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism*; D009599:Nitroprusside / Q000494:pharmacology*; D010761:Phosphorus Radioisotopes; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013379:Substrate Specificity
D007527:Isoenzymes; D010761:Phosphorus Radioisotopes; D009243:NAD; D009599:Nitroprusside; D000246:Adenosine Diphosphate Ribose; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D000444:Aldehyde Dehydrogenase; D004229:Dithiothreitol