The copper- and zinc-containing superoxide dismutase from Escherichia coli: molecular weight and stability.
The periplasmic Cu,Zn superoxide dismutase (Cu,ZnSOD) from Escherichia coli has been shown by sedimentation equilibrium to be a monomer with a molecular weight of approximately 17,000. The enzyme suffered a reversible inactivation when heated to 70 degrees C. This was minimized by added Cu(II) or Zn(II). Heat lability was greater in phosphate than in Tris buffer. The enzyme exhibited a time-dependent inactivation by Hg(II) and this too was greater in phosphate than in Tris. This behavior can be explained by a modest affinity of the enzyme for Cu(II) and Zn(II) which results in a dissociation/association equilibrium. Elevation of the temperature shifts this equilibrium toward dissociation and phosphate sequesters the released metals making them less available for reinsertion at the active site. Hg(II) competes for occupancy of the active site and there were more unoccupied sites in phosphate than in Tris. A parallel was drawn between the E. coli Cu,ZnSOD and FALS varients of human Cu,ZnSOD, which are also relatively unstable and exhibit low affinity for Cu(II).
Benov
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Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710, USA.
Sage
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Fridovich
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D001386:Azides / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry; D002413:Cations, Divalent; D003300:Copper / Q000494:pharmacology; D003486:Cyanides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D006358:Hot Temperature; D008628:Mercury / Q000494:pharmacology; D008970:Molecular Weight; D010710:Phosphates / Q000494:pharmacology; D011489:Protein Denaturation / Q000187:drug effects; D013482:Superoxide Dismutase / Q000737:chemistry*; D014325:Tromethamine / Q000494:pharmacology; D014461:Ultracentrifugation
D001386:Azides; D001426:Bacterial Proteins; D002413:Cations, Divalent; D003486:Cyanides; D004791:Enzyme Inhibitors; D010710:Phosphates; D014325:Tromethamine; D003300:Copper; D013482:Superoxide Dismutase; D008628:Mercury