Proton NMR study of the heme environment in bacterial quinol oxidases

Jie Zhang, Jeffrey P. Osborne, Robert B. Gennis, Xiaotang Wang

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The heme environment and ligand binding properties of two relatively large membrane proteins containing multiple paramagnetic metal centers, cytochrome bo3 and bd quinol oxidases, have been studied by high field proton nuclear magnetic resonance (NMR) spectroscopy. The oxidized bo3 enzyme displays well-resolved hyperfine-shifted 1H NMR resonance assignable to the low-spin heme b center. The observed spectral changes induced by addition of cyanide to the protein were attributed to the structural perturbations on the low-spin heme (heme b) center by cyanide ligation to the nearby high-spin heme (heme o) of the protein. The oxidized bd oxidase shows extremely broad signals in the spectral region where protons near high-spin heme centers resonate. Addition of cyanide to the oxidized bd enzyme induced no detectable perturbations on the observed hyperfine signals, indicating the insensitive nature of this heme center toward cyanide. The proton signals near the low-spin heme b558 center are only observed in the presence of 20% formamide, consistent with a critical role of viscosity in detecting NMR signals of large membrane proteins. The reduced bd protein also displays hyperfine-shifted 1H NMR signals, indicating that the high-spin heme centers (hemes b595 and d) remain high-spin upon chemical reduction. The results presented here demonstrate that structural changes of one metal center can significantly influence the structural properties of other nearby metal center(s) in large membrane paramagnetic metalloproteins.

Original languageEnglish (US)
Pages (from-to)186-191
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume421
Issue number2
DOIs
StatePublished - Jan 15 2004
Externally publishedYes

Fingerprint

Heme
Protons
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Cyanides
Metals
Membrane Proteins
duroquinol oxidase
Metalloproteins
Proteins
Enzymes
Cytochromes
Viscosity
Nuclear magnetic resonance spectroscopy
Ligation
Structural properties
Oxidoreductases
Display devices
Ligands
Membranes

Keywords

  • Heme center
  • Ligand binding
  • Nuclear magnetic resonance
  • Quinol oxidase
  • Spin state
  • Structural perturbation

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Proton NMR study of the heme environment in bacterial quinol oxidases. / Zhang, Jie; Osborne, Jeffrey P.; Gennis, Robert B.; Wang, Xiaotang.

In: Archives of Biochemistry and Biophysics, Vol. 421, No. 2, 15.01.2004, p. 186-191.

Research output: Contribution to journalArticle

Zhang, Jie ; Osborne, Jeffrey P. ; Gennis, Robert B. ; Wang, Xiaotang. / Proton NMR study of the heme environment in bacterial quinol oxidases. In: Archives of Biochemistry and Biophysics. 2004 ; Vol. 421, No. 2. pp. 186-191.
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