title: In vivo competition between iron and manganese for occupancy of the active site region of the manganese-superoxide dismutase of Escherichia coli.
abstract: Three forms of the dimeric manganese superoxide dismutase (MnSOD) were isolated from aerobically grown Escherichia coli which contained 2 Mn, 1 Mn and 1 Fe, or 2 Fe, respectively. These are designated Mn2-MnSOD, Mn,Fe-MnSOD, and Fe2-MnSOD. Substitution of iron in place of manganese, eliminated catalytic activity, decreased the isoelectric point, and increased the native electrophoretic anodic mobility, although circular dichroism, high performance liquid chromatography gel exclusion chromatography, and sedimentation equilibrium revealed no gross changes in conformation. Moreover, replacement of iron by manganese restored enzymatic activity. Fe2-MnSOD and the iron-superoxide (FeSOD) of E. coli exhibit distinct optical absorption spectra. These data indicate that the active site environments of E. coli MnSOD and FeSOD must differ. They also indicate that competition between iron and manganese for nascent MnSOD polypeptide chains occurs in vivo, and copurification of these variably substituted MnSODs can explain the substoichiometric manganese contents and the variable specific activities which have been reported for this enzyme.
authors.affiliation: Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710.
authors.forename: W F
authors.initials: WF
authors.lastname: Beyer
authors.forename: I
authors.initials: I
authors.lastname: Fridovich
mesh_terms: D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development; D007501:Iron / Q000378:metabolism*; D007700:Kinetics; D008345:Manganese / Q000378:metabolism*; D008970:Molecular Weight; D011487:Protein Conformation; D013053:Spectrophotometry; D013482:Superoxide Dismutase / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D008345:Manganese; D007501:Iron; D013482:Superoxide Dismutase