Carbonic anhydrase III. Oxidative modification in vivo and loss of phosphatase activity during aging.
Oxidative modification of DNA, lipids, and proteins occurs as a consequence of reaction with free radicals and activated oxygen. Oxidative modification of total cellular proteins has been described under many pathologic and experimental conditions, but no specific proteins have been identified as in vivo targets for oxidative modification. Utilizing an immunochemical method for detection of oxidatively modified proteins, we identified a protein in rat liver that was highly oxidized. It was purified to homogeneity and identified as carbonic anhydrase, isozyme III. Its characteristics match those previously described for a protein that was lost during aging of the rat, senescence marker protein-1. Carbonic anhydrase III was purified from rats aged 2, 10, and 18 months, and the proteins were characterized. All three preparations were highly oxidatively modified as assessed by their carbonyl content. The enzyme has three known catalytic activities, and the specific activities for carbon dioxide hydration and for ester hydrolysis decreased during aging by approximately 30%. However, the third activity, that of a phosphatase, was virtually lost during aging. While the physiologic role of carbonic anhydrase III is unknown, we suggest that it functions in an oxidizing environment, which leads to its own oxidative modification.
Cabiscol
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Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892-0320, USA.
Levine
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RL
D000375:Aging / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002256:Carbonic Anhydrases / Q000302:isolation & purification / Q000378:metabolism*; D002384:Catalysis; D005978:Glutathione / Q000378:metabolism; D006358:Hot Temperature; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D011489:Protein Denaturation; D051381:Rats; D011916:Rats, Inbred F344
D010744:Phosphoric Monoester Hydrolases; D002256:Carbonic Anhydrases; D005978:Glutathione