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Manganese superoxide dismutase fromThermus thermophilus:A structural model refined at 1.8 Å resolution

Authors
Journal
Journal of Molecular Biology
0022-2836
Publisher
Elsevier
Publication Date
Volume
219
Issue
2
Identifiers
DOI: 10.1016/0022-2836(91)90569-r
Keywords
  • Superoxide
  • Superoxide Dismutase
  • Refinement
  • Manganese(Iii)
  • Manganese(Ii)
  • Dimer
  • Tetramer
  • Domain Interface
Disciplines
  • Biology
  • Mathematics

Abstract

Abstract The structure of Mn(III) superoxide dismutase (Mn(III)SOD) from Thermus thermophilus, a tetramer of chains 203 residues in length, has been refined by restrained least-squares methods. The R-factor ( = ∑¦¦F o ¦− ¦F c ¦¦ ∑¦F o ¦ ) for the 54,056 unique reflections measured between 10·0 and 1·8 Å (96% of all possible reflections) is 0·176 for a model comprising the protein dimer and 180 bound solvents, the asymmetric unit of the P4 12 12 cell. The monomer chain forms two domains as determined by distance plots: the N-terminal domain is dominated by two long antiparallel helices (residues 21 to 45 and 69 to 89) and the C-terminal domain (residues 100 to 203) is an α + β structure including a three-stranded sheet. Features that may be important for the folding and function of this MnSOD include: (1) a cis-proline in a turn preceding the first long helix; (2) a residue inserted at position 30 that distorts the helix near the first Mn ligand; and (3) the locations of glycine and proline residues in the domain connector (residues 92 to 99) and in the vicinity of the short cross connection (residues 150 to 159) that links two strands of the β-sheet. Domain-domain contacts include salt bridges between arginine residues and acidic side chains, an extensive hydrophobic interface, and at least ten hydrogen-bonded interactions. The tetramer possesses 222 symmetry but is held together by only two types of interfaces. The dimer interface at the non-crystallographic dyad is extensive (1000 Å 2 buried surface/ monomer) and incorporates 17 trapped or structural solvents. The dimer interface at the crystallographic dyad buries fewer residues (750 Å 2/monomer) and resembles a snap fastener in which a type I turn thrusts into a hydrophobic basket formed by a ring of helices in the opposing chain. Each of the metal sites is fully occupied, with the Mn(III) five-co-ordinate in trigonal bipyramidal geometry. One of the axial ligands is solvent; the four protein ligands are His28, His83, Asp166 and His170. Surrounding the metal-ligand cluster is a shell of predominantly hydrophobic residues from both chains of the asymmetric unit (Phe86A, Trp87A, Trp132A, Trp168A, Tyr183A, Tyr172B, Tyr173B), and both chains collaborate in the formation of a solvent-lined channel that terminates at Tyr36 and His32 near the metal ion and is presumed to be the path by which substrate or other inner-sphere ligands reach the metal. A pocket adjoining the metal, formed by His33, Trp87, His83 and Tyr36, is postulated to be the substrate-binding site. Refinement of 2.3 Å data from crystals reduced with dithionite indicates that the co-ordination geometry at the metal is not changed by reduction.

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