Measuring distances in proteins by saturation-recovery EPR.

Donald J. Hirsh, Gary W Brudvig

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We describe a protocol for detecting electron spin-spin interactions between a radical and a metal ion in a protein or protein complex by saturation-recovery electron paramagnetic resonance (EPR). This protocol can be used with a protein containing an endogenous metal center and either an endogenous or synthetic radical species. We suggest a two-step approach whereby dipole-dipole or exchange interactions are first detected by continuous-wave EPR experiments and then quantified by saturation-recovery EPR. The latter measurements make it possible to measure long distances to within a few Angstroms. The protocol for making distance measurements by saturation-recovery EPR will take approximately 6 days to complete.

Original languageEnglish
Pages (from-to)1770-1781
Number of pages12
JournalNature Protocols
Volume2
Issue number7
DOIs
Publication statusPublished - 2007

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Electron Spin Resonance Spectroscopy
Paramagnetic resonance
Recovery
Proteins
Metals
Distance measurement
Exchange interactions
Metal ions
Electrons
Ions
Experiments

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Measuring distances in proteins by saturation-recovery EPR. / Hirsh, Donald J.; Brudvig, Gary W.

In: Nature Protocols, Vol. 2, No. 7, 2007, p. 1770-1781.

Research output: Contribution to journalArticle

Hirsh, Donald J. ; Brudvig, Gary W. / Measuring distances in proteins by saturation-recovery EPR. In: Nature Protocols. 2007 ; Vol. 2, No. 7. pp. 1770-1781.
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