A Simple Proposal That Can Explain the Inactivity of Metal-Substituted Superoxide Dismutases

• Published in: Journal of the American Chemical Society 1998-01, Vol.120 (3), p.461-467
• Main Authors: Vance, C. K, Miller, A.-F
• Format: Article
• Language: English
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/ja972060j

We propose that the apparent catalytic inactivity of Mn- and Fe-substituted superoxide dismutases (SODs) reflects E°s that are either lower (Fe-sub-(Mn)SOD) or higher (Mn-sub-(Fe)SOD) than those of native Fe- or Mn-SODs. In support, we show that the E° of Fe-sub-(Mn)SOD (Fe substituted into Mn-SOD protein) is −240 mV vs NHE, almost 0.5 V lower than our E° of 220 mV for Fe-SOD. The E° of Fe-sub-(Mn)SOD is lower than that of O2/O2 •- and therefore is sufficient to explain Fe-sub-(Mn)SOD's inactivity. Indeed, Fe-sub-(Mn)SOD is shown to be unable to oxidize O2 •-. Alternate causes of inactivity are ruled out by our demonstration that Fe-sub-(Mn)SOD retains the ability to reduce O2 •-. Thus, the active site remains active with respect to substrate binding and proton and electron transfer. Finally, we show that Fe-sub-(Mn)SOD's inactivity with respect to O2 •- oxidation cannot be solely due to competitive inhibition by OH-. Thus, our proposal provides a simple chemical basis for the observed catalytic inactivity of metal-exchanged Mn- or Fe-SODs and suggests that these strongly homologous enzymes may provide important insights into mechanisms of redox midpoint potential tuning in proteins.