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Bicarbonate Enhances the Peroxidase Activity of Cu,Zn-Superoxide Dismutase

Journal of Biological Chemistry, ISSN: 0021-9258, Vol: 274, Issue: 40, Page: 28233-28239
1999
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We examined the effect of bicarbonate on the peroxidase activity of copper-zinc superoxide dismutase (SOD1), using the nitrite anion as a peroxidase probe. Oxidation of nitrite by the enzyme-bound oxidant results in the formation of the nitrogen dioxide radical, which was measured by monitoring 5-nitro-γ-tocopherol formation. Results indicate that the presence of bicarbonate is not required for the peroxidase activity of SOD1, as monitored by the SOD1/H 2 O 2 -mediated nitration of γ-tocopherol in the presence of nitrite. However, bicarbonate enhanced SOD1/H 2 O 2 -dependent oxidation of tocopherols in the presence and absence of nitrite and dramatically enhanced SOD1/H 2 O 2 -mediated oxidation of unsaturated lipid in the presence of nitrite. These results, coupled with the finding that bicarbonate protects against inactivation of SOD1 by H 2 O 2, suggest that SOD1/H 2 O 2 oxidizes the bicarbonate anion to the carbonate radical anion. Thus, the amplification of peroxidase activity of SOD1/H 2 O 2 by bicarbonate is attributed to the intermediary role of the diffusible oxidant, the carbonate radical anion. We conclude that, contrary to a previous report (Sankarapandi, S., and Zweier, J. L. (1999) J. Biol. Chem. 274, 1226–1232), bicarbonate is not required for peroxidase activity mediated by SOD1 and H 2 O 2. However, bicarbonate enhanced the peroxidase activity of SOD1 via formation of a putative carbonate radical anion. Biological implications of the carbonate radical anion in free radical biology are discussed.

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