Carbon dioxide stimulates peroxynitrite-mediated nitration of tyrosine residues and inhibits oxidation of methionine residues of glutamine synthetase: both modifications mimic effects of adenylylation.
The activity of glutamine synthetase (EC 6.3.1.2) from Escherichia coli is regulated by the cyclic adenylylation and deadenylylation of Tyr-397 in each of the enzyme's 12 identical subunits. The nitration of Tyr-397 or of the nearby Tyr-326 by peroxynitrite can convert the unadenylylated enzyme to a form exhibiting regulatory characteristics similar to the form obtained by adenylylation. The adenylylated conformation can also be elicited by the oxidation of surface-exposed methionine residues to methionine sulfoxide. However, the nitration of tyrosine residues and the oxidation of methionine residues are oppositely directed by the presence and absence of CO2. At physiological concentrations of CO2, pH 7.4, nitration occurs but oxidation of methionine residues is inhibited. Conversely, in the absence of CO2 methionine oxidation is stimulated and nitration of tyrosine is prevented. It was further established that adenylylation of Tyr-397 precludes its nitration by peroxynitrite. Furthermore, nitration of Tyr-326 together with either nitration or adenylylation of Tyr-397 leads to inactivation of the enzyme. These results demonstrate that CO2 can alter the course of peroxynitrite-dependent reactions and serve notice that (i) the reactions have physiological significance only if they are shown to occur at physiological concentrations of CO2 and physiological pH; and (ii) the peroxynitrite-dependent nitration of tyrosine residues or the oxidation of methionine residues of metabolically regulated proteins can seriously compromise their biological function.
Berlett
B S
BS
Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892-0342, USA.
Levine
R L
RL
Stadtman
E R
ER
D000249:Adenosine Monophosphate; D002245:Carbon Dioxide / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D005974:Glutamate-Ammonia Ligase / Q000187:drug effects / Q000378:metabolism*; D008715:Methionine; D008956:Models, Chemical; D008958:Models, Molecular; D009566:Nitrates / Q000494:pharmacology; D010084:Oxidation-Reduction; D014443:Tyrosine
D009566:Nitrates; D002245:Carbon Dioxide; C039642:peroxynitric acid; D000249:Adenosine Monophosphate; D014443:Tyrosine; D008715:Methionine; D005974:Glutamate-Ammonia Ligase