title: Geometric and electronic structures of manganese-substituted iron superoxide dismutase.
abstract: The active-site structures of the oxidized and reduced forms of manganese-substituted iron superoxide dismutase (Mn(Fe)SOD) are examined, for the first time, using a combination of spectroscopic and computational methods. On the basis of electronic absorption, circular dichroism (CD), magnetic CD (MCD), and variable-temperature variable-field MCD data obtained for oxidized Mn(Fe)SOD, we propose that the active site of this species is virtually identical to that of wild-type manganese SOD (MnSOD), with both containing a metal ion that resides in a trigonal bipyramidal ligand environment. This proposal is corroborated by quantum mechanical/molecular mechanical (QM/MM) computations performed on complete protein models of Mn(Fe)SOD in both its oxidized and reduced states and, for comparison, wild-type (WT) MnSOD. The major differences between the QM/MM optimized active sites of WT MnSOD and Mn(Fe)SOD are a smaller (His)N-Mn-N(His) equatorial angle and a longer (Gln146(69))NH···O(sol) H-bond distance in the metal-substituted protein. Importantly, these modest geometric differences are consistent with our spectroscopic data obtained for the oxidized proteins and high-field electron paramagnetic resonance spectra reported previously for reduced Mn(Fe)SOD and MnSOD. As Mn(Fe)SOD exhibits a reduction midpoint potential (E(m)) almost 700 mV higher than that of MnSOD, which has been shown to be sufficient for explaining the lack of SOD activity displayed by the metal-subtituted species (Vance, C. K.; Miller, A. F. Biochemistry 2001, 40, 13079-13087), E(m)'s were computed for our experimentally validated QM/MM optimized models of Mn(Fe)SOD and MnSOD. These computations properly reproduce the experimental trend and reveal that the drastically elevated E(m) of the metal substituted protein stems from a larger separation between the second-sphere Gln residue and the coordinated solvent in Mn(Fe)SOD relative to MnSOD, which causes a weakening of the corresponding H-bond interaction in the oxidized state and alleviates steric crowding in the reduced state.
authors: Department of Chemistry, University of Wisconsin-Madison, Madison Wisconsin 53706, USA.
authors: Timothy A
authors: TA
authors: Jackson
authors: Craig T
authors: CT
authors: Gutman
authors: James
authors: J
authors: Maliekal
authors: Anne-Frances
authors: AF
authors: Miller
authors: Thomas C
authors: TC
authors: Brunold
mesh_terms: D020134:Catalytic Domain; D004583:Electrons; D008345:Manganese / Q000378:metabolism*; D008958:Models, Molecular; D010084:Oxidation-Reduction; D013057:Spectrum Analysis; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D008345:Manganese; D013482:Superoxide Dismutase