Q-band electron nuclear double resonance (ENDOR) and X-band EPR of the sulfobetaine 12 heat-treated cytochrome c oxidase complex

Siegfried M. Musser, Yang Cheng Fann, Ryszard J. Gurbiel, Brian M. Hoffman, Sunney I. Chan*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Heat treatment of the bovine cytochrome c oxidase complex in the zwitterionic detergent sulfobetaine 12 (SB-12) results in loss of subunit III and the appearance of a type II copper center as characterized by electron paramagnetic resonance (EPR) spectroscopy. Previous authors (Nilsson, T., Copeland, R. A., Smith, P. A., and Chan, S. I. (1988) Biochemistry 27, 8254- 8260) have interpreted this type II copper center as a modified version of the Cu(A) site. By using electron nuclear double resonance spectroscopy, it is found that the Cu(A) proton and nitrogen resonances remain present in the SB-12 heat-treated enzyme and that three new nitrogen resonances appear having hyperfine coupling constants consistent with histidine ligation. These hyperfine coupling constants correlate well with those recently found for the Cu(B) histidines from the cytochrome aa 3-600 quinol oxidase from Bacillus subtilis (Fann, Y. C., Ahmed, I., Blackburn, N. J., Boswell, J. S., Verkhovskaya, M. L., Hoffman, B. M., and Wikstrom, M. (1995) Biochemistry 34, 10245-10255). In addition, the total EPR-detectable copper concentration per enzyme molecule approximately doubles upon SB-12 heat treatment. Finally, the observed type II copper EPR spectrum is virtually indistinguishable from the EPR spectrum of Cu(B) of the as-isolated cytochrome bo 3 complex from Escherichia coli. These data indicate that the type II copper species that appears results from a breaking of the strong antiferromagnetic coupling of the heme a 3-Cu(B) binuclear center.

Original languageEnglish (US)
Pages (from-to)203-209
Number of pages7
JournalJournal of Biological Chemistry
Issue number1
StatePublished - 1997

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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