1EN6

CRYSTAL STRUCTURE ANALYSIS OF THE E. COLI MANGANESE SUPEROXIDE DISMUTASE Q146L MUTANT

1EN6 の概要

• エントリーDOI: 10.2210/pdb1en6/pdb
• 関連するPDBエントリー: 1EN4 1EN5 1MMM 1VEW
• 分子名称: MANGANESE SUPEROXIDE DISMUTASE, MANGANESE (II) ION (3 entities in total)
• 機能のキーワード: proton shuttle, q146l, mutant, manganese superoxide dismutase, oxidoreductase
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 92147.38

構造登録者

Edwards, R.A.,Whittaker, M.M.,Baker, E.N.,Whittaker, J.W.,Jameson, G.B. (登録日: 2000-03-20, 公開日: 2001-06-27, 最終更新日: 2024-02-07)

主引用文献

Edwards, R.A.,Whittaker, M.M.,Whittaker, J.W.,Baker, E.N.,Jameson, G.B.
Outer sphere mutations perturb metal reactivity in manganese superoxide dismutase.
Biochemistry, 40:15-27, 2001

PubMed Abstract: Tyrosine 34 and glutamine 146 are highly conserved outer sphere residues in the mononuclear manganese active site of Escherichia coli manganese superoxide dismutase. Biochemical and spectroscopic characterization of site-directed mutants has allowed functional characterization of these residues in the wild-type (wt) enzyme. X-ray crystallographic analysis of three mutants (Y34F, Q146L, and Q146H) reveal subtle changes in the protein structures. The Y34A mutant, as well as the previously reported Y34F mutant, retained essentially the full superoxide dismutase activity of the wild-type enzyme, and the X-ray crystal structure of Y34F manganese superoxide dismutase shows that mutation of this strictly conserved residue has only minor effects on the positions of active site residues and the organized water in the substrate access funnel. Mutation of the outer sphere solvent pocket residue Q146 has more dramatic effects. The Q146E mutant is isolated as an apoprotein lacking dismutase activity. Q146L and Q146H mutants retain only 5-10% of the dismutase activity of the wild-type enzyme. The absorption and circular dichroism spectra of the Q146H mutant resemble corresponding data for the superoxide dismutase from a hyperthermophilic archaeon, Pyrobaculum aerophilum, which is active in both Mn and Fe forms. Interestingly, the iron-substituted Q146H protein also exhibits low dismutase activity, which increases at lower pH. Mutation of glutamine 146 disrupts the hydrogen-bonding network in the active site and has a greater effect on protein structure than does the Y34F mutant, with rearrangement of the tyrosine 34 and tryptophan 128 side chains.

PubMed: 11141052
DOI: 10.1021/bi0018943

実験手法

X-RAY DIFFRACTION (2 Å)