Oxidation of Neurospora crassa glutamine synthetase.
The glutamine synthetase of Neurospora crassa, either purified or in cell extracts, was inactivated by ascorbate plus FeCl3 and by H2O2 plus FeSO4. The inactivation reaction was oxygen dependent, inhibited by MnCl2 and EDTA, and stimulated in cell extracts by sodium azide. This inactivation could also be brought about by adding NADPH to the cell extract. The alpha and beta polypeptides of the active glutamine synthetase were modified by these inactivating reactions, giving rise to two novel acidic polypeptides. These modifications were observed with the purified enzyme, with cell extracts, and under in vivo conditions in which glutamine synthetase is degraded. The modified glutamine synthetase was more susceptible to endogenous phenylmethylsulfonyl fluoride-insensitive proteolytic activity, which was inhibited by MnCl2 and stimulated by EDTA. The possible physiological relevance of enzyme oxidation is discussed.
Aguirre
J
J
Hansberg
W
W
D001205:Ascorbic Acid / Q000494:pharmacology; D002712:Chlorides; D004492:Edetic Acid / Q000494:pharmacology; D005296:Ferrous Compounds / Q000494:pharmacology; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D046911:Macromolecular Substances; D008345:Manganese / Q000494:pharmacology; D017895:Manganese Compounds; D009249:NADP / Q000378:metabolism; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D010084:Oxidation-Reduction; D010447:Peptide Hydrolases / Q000378:metabolism
D002712:Chlorides; D005296:Ferrous Compounds; D046911:Macromolecular Substances; D017895:Manganese Compounds; C020748:ferrous sulfate; D008345:Manganese; D009249:NADP; D004492:Edetic Acid; D006861:Hydrogen Peroxide; D010447:Peptide Hydrolases; D005974:Glutamate-Ammonia Ligase; D001205:Ascorbic Acid; C025340:manganese chloride; C029451:ferrous chloride