Electron paramagnetic resonance studies of succinate:ubiquinone oxidoreductase from Paracoccus denitrificans: Evidence for a magnetic interaction between the 3Fe-4S cluster and cytochrome b

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Abstract

Electron paramagnetic resonance (EPR) studies of succinate:ubiquinone oxidoreductase (SQR) from Paracoccus denitrificans have been undertaken in the purified and membrane-bound states, Spectroscopic ''signatures'' accounting for the three iron-sulfur clusters (2Fe-2S, 3Fe-4S, and 4Fe-4S), cytochrome b, flavin, and protein-bound ubisemiquinone radicals have been obtained in air-oxidized, succinate-reduced, and dithionite-reduced preparations at 4-10 K. Spectra obtained at 170 K in the presence of excess succinate showed a signal typical of that of a flavin radical, but superimposed with another signal. The superimposed signal originated from two bound ubisemiquinones, as shown by spectral simulations, Power saturation measurements performed on the air-oxidized enzyme provided evidence for a weak magnetic dipolar interaction operating between the oxidized 3Fe-4S cluster and the oxidized cytochrome b. Power saturation experiments performed on the succinate- and dithionite-reduced forms of the enzyme demonstrated that the 4Fe-4S cluster is coupled weakly to both the 2Fe-2S and the 3Fe-4S clusters, Quantitative interpretation of these power saturation experiments has been achieved through redox calculations. They revealed that a spin-spin interaction between the reduced 3Fe-4S cluster and the cytochrome b (oxidized) may also exist. These findings form the first direct EPR evidence for a close proximity (less than or equal to 2 nm) of the high potential 3Fe-4S cluster, situated in the succinate dehydrogenase part of the enzyme, and the low potential, low spin b-heme in the membrane anchor of the enzyme.

Details

Authors
  • A. Reginald Waldeck
  • Michael H. B. Stowell
  • Hung Kay Lee
  • Shao-Ching Hung
  • Mikael Matsson
  • Lars Hederstedt
  • Brian A.C. Ackrell
  • Sunney I. Chan
Organisations
External organisations
  • California Institute of Technology
  • University of California, San Francisco
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Microbiology
Original languageEnglish
JournalJournal of Biological Chemistry
Publication statusPublished - 1997
Publication categoryResearch
Peer-reviewedYes