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

doi:10.1016/0022-2836(91)90569-R

Journal of Molecular Biology

Volume 219, Issue 2, 20 May 1991, Pages 335-358

https://doi.org/10.1016/0022-2836(91)90569-R Get rights and content

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 P4₁2₁2 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 Å² buried surface/ monomer) and incorporates 17 trapped or structural solvents. The dimer interface at the crystallographic dyad buries fewer residues (750 Å²/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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^☆: This research was supported by grants from the National Institutes of Health (GM16429) to M.L.L. and to Dr J. A. Fee (GM35189). Diffraction experiments were conducted at the Multiwire Area Detector Facility at the University of California, San Diego, and some of the computations were carried out at the San Diego Supercomputer Center.

^‡: Present address: Monsanto Corporate Research, Mailzone BB4K, 700 Chesterfield Village. St Louis, MO 63198, U.S.A.

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Journal of Molecular Biology

ArticleManganese superoxide dismutase from Thermus thermophilus: A structural model refined at 1.8 Å resolution☆

Abstract

J. Biol. Chem.

J. Biol. Chem.

Arch. Biochem. Biophys.

J. Biol. Chem.

Arch. Biochem. Biophys.

J. Mol. Biol.

Advan. Protein Chem.

J. Mol. Graph.

J. Biol. Chem.

Arch. Biochem. Biophys.

J. Biol. Chem.

Inorg. Nucl. Chem. Letters

J. Biol. Chem.

J. Biol. Chem.

J. Mol. Biol.

J. Mol. Biol.

Methods Enzymol.

FEBS Letters

FEBS Letters

J. Mol. Biol.

J. Mol. Biol.

J. Mol. Biol.

Biochim. Biophys. Acta

J. Mol. Biol.

Methods Enzymol.

Methods Enzymol.

Advan. Protein Chem.

Biochim. Biophys. Acta

FEBS Letters

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Advan. Inorg. Chem.

J. Mol. Biol.

J. Mol. Biol.

J. Biol. Chem.

Ionic strength dependence of enzyme-substrate interactions. Monte-Carlo and Poisson-Boltzmann results for superoxide dismutase

J. Phys. Chem.

Synthetic model approach to the manganese(III) acid phosphatase and its iron(III)-substituted form

J. Amer. Chem. Soc.

Efficient production of active human manganese superoxide dismutase in Escherichia coli

Biotechnology

Evidence of the breaking of the copper imidazolate bridge in copper/cobalt substituted superoxide dismutase upon reduction of the copper(TI) centers

J. Amer. Chem. Soc.

Deciphering the message in protein sequences: tolerance to amino acid substitutions

Science

Superoxide dismutase from Bacillus stearothermophilus. Complete amino acid sequence of a manganese enzyme

Biochemistry

Steady-state kinetic studies of superoxide dismutases: properties of the iron containing protein from Escherichia coli

J. Amer. Chem. Soc.

Kinetic studies of superoxide dismutases: properties of the manganese containing protein from Thermus thermophilus

J. Amer. Chem. Soc.

Article
Manganese superoxide dismutase from Thermus thermophilus: A structural model refined at 1.8 Å resolution☆