Calometric studies on the tight binding metal interactions of Escherichia coli manganese superoxide dismutase

Kazunori Mizuno, Mei M. Whittaker, Hans Peter Bächinger, James Whittaker

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Escherichia coli apomanganese superoxide dismutase, prepared by removing the native metal ion under denaturing conditions, exhibits thermally triggered metal uptake behavior previously observed for thermophilic and hyperthermophilic superoxide dismutases but over a lower temperature range. Differential scanning calorimetry of aposuperoxide dismutase and metalated superoxide dismutase unfolding transitions has provided quantitative estimates of the metal binding affinities for manganese superoxide dismutase. The binding constant for Mn(II) (KMn(II) = 3.2 × 108 M-1) is surprisingly low in light of the essentially irreversible metal binding characteristic of this family of proteins and indicates that metal binding and release processes are dominated by kinetic, rather than thermodynamic, constraints. The kinetic stability of the metalloprotein complex can be traced to stabilization by elements of the protein that are independent of the presence or absence of the metal ion reflected in the thermally triggered metalation characteristic of these proteins. Binding constants for Mn(III), Fe(II), and Fe(III) complexes were estimated using quasireversible values for the unfolding enthalpy and ΔCp for apo-Mn superoxide dismutase and the observed T m values for unfolding the metalated species in the absence of denaturants. For manganese and iron complexes, an oxidation state-dependent binding affinity reflects the protein perturbation of the metal redox potential.

Original languageEnglish (US)
Pages (from-to)27339-27344
Number of pages6
JournalJournal of Biological Chemistry
Volume279
Issue number26
DOIs
StatePublished - Jun 25 2004

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Escherichia coli
Superoxide Dismutase
Metals
Metal ions
Proteins
Metalloproteins
Kinetics
Manganese
Ions
Differential scanning calorimetry
Enthalpy
Differential Scanning Calorimetry
Iron
Stabilization
Thermodynamics
Oxidation-Reduction
Oxidation
Temperature

ASJC Scopus subject areas

  • Biochemistry

Cite this

Calometric studies on the tight binding metal interactions of Escherichia coli manganese superoxide dismutase. / Mizuno, Kazunori; Whittaker, Mei M.; Bächinger, Hans Peter; Whittaker, James.

In: Journal of Biological Chemistry, Vol. 279, No. 26, 25.06.2004, p. 27339-27344.

Research output: Contribution to journalArticle

Mizuno, Kazunori ; Whittaker, Mei M. ; Bächinger, Hans Peter ; Whittaker, James. / Calometric studies on the tight binding metal interactions of Escherichia coli manganese superoxide dismutase. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 26. pp. 27339-27344.
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