Nitroxide spin labels as EPR reporters of the relaxation and magnetic properties of the heme-copper site in cytochrome bo 3, E. coli

Vasily S. Oganesyan, Gaye F. White, Sarah Field, Sophie Marritt, Robert B. Gennis, Lai Lai Yap, Andrew J. Thomson

Research output: Contribution to journalArticle

Abstract

A nitroxide spin label (SL) has been used to probe the electron spin relaxation times and the magnetic states of the oxygen-binding heme-copper dinuclear site in Escherichia coli cytochrome bo 3, a quinol oxidase (QO), in different oxidation states. The spin lattice relaxation times, T 1, of the SL are enhanced by the paramagnetic metal sites in QO and hence show a strong dependence on the oxidation state of the latter. A new, general form of equations and a computer simulation program have been developed for the calculation of relaxation enhancement by an arbitrary fast relaxing spin system of S ≤ 1/2. This has allowed us to obtain an accurate estimate of the transverse relaxation time, T 2, of the dinuclear coupled pair Fe(III)-CuB(II) in the oxidized form of QO that is too short to measure directly. In the case of the F state, the relaxation properties of the heme-copper center have been shown to be consistent with a ferryl [Fe(IV)=O] heme and CuB(II) coupled by approximately 1.5-3 cm-1 to a radical. The magnitude suggests that the coupling arises from a radical form of the covalently linked tyrosine-histidine ligand to Cu(II) with unpaired spin density primarily on the tyrosine component. This work demonstrates that nitroxide SLs are potentially valuable tools to probe both the relaxation and the magnetic properties of multinuclear high-spin paramagnetic active sites in proteins that are otherwise not accessible from direct EPR measurements.

Original languageEnglish (US)
Pages (from-to)1255-1264
Number of pages10
JournalJournal of Biological Inorganic Chemistry
Volume15
Issue number8
DOIs
StatePublished - Nov 1 2010

Fingerprint

Spin Labels
Heme
Relaxation time
Paramagnetic resonance
Copper
Magnetic properties
Tyrosine
Cytochromes a3
Oxidation
Spin-lattice relaxation
Cytochromes
Histidine
Computer Simulation
Escherichia coli
Catalytic Domain
Software
Metals
Electrons
Oxygen
Ligands

Keywords

  • Electron paramagnetic resonance
  • Metalloprotein
  • Relaxation time
  • Spin labels
  • Spin relaxation enhancement

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry

Cite this

Nitroxide spin labels as EPR reporters of the relaxation and magnetic properties of the heme-copper site in cytochrome bo 3, E. coli. / Oganesyan, Vasily S.; White, Gaye F.; Field, Sarah; Marritt, Sophie; Gennis, Robert B.; Yap, Lai Lai; Thomson, Andrew J.

In: Journal of Biological Inorganic Chemistry, Vol. 15, No. 8, 01.11.2010, p. 1255-1264.

Research output: Contribution to journalArticle

Oganesyan, Vasily S. ; White, Gaye F. ; Field, Sarah ; Marritt, Sophie ; Gennis, Robert B. ; Yap, Lai Lai ; Thomson, Andrew J. / Nitroxide spin labels as EPR reporters of the relaxation and magnetic properties of the heme-copper site in cytochrome bo 3, E. coli. In: Journal of Biological Inorganic Chemistry. 2010 ; Vol. 15, No. 8. pp. 1255-1264.
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AU - Thomson, Andrew J.

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AB - A nitroxide spin label (SL) has been used to probe the electron spin relaxation times and the magnetic states of the oxygen-binding heme-copper dinuclear site in Escherichia coli cytochrome bo 3, a quinol oxidase (QO), in different oxidation states. The spin lattice relaxation times, T 1, of the SL are enhanced by the paramagnetic metal sites in QO and hence show a strong dependence on the oxidation state of the latter. A new, general form of equations and a computer simulation program have been developed for the calculation of relaxation enhancement by an arbitrary fast relaxing spin system of S ≤ 1/2. This has allowed us to obtain an accurate estimate of the transverse relaxation time, T 2, of the dinuclear coupled pair Fe(III)-CuB(II) in the oxidized form of QO that is too short to measure directly. In the case of the F state, the relaxation properties of the heme-copper center have been shown to be consistent with a ferryl [Fe(IV)=O] heme and CuB(II) coupled by approximately 1.5-3 cm-1 to a radical. The magnitude suggests that the coupling arises from a radical form of the covalently linked tyrosine-histidine ligand to Cu(II) with unpaired spin density primarily on the tyrosine component. This work demonstrates that nitroxide SLs are potentially valuable tools to probe both the relaxation and the magnetic properties of multinuclear high-spin paramagnetic active sites in proteins that are otherwise not accessible from direct EPR measurements.

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