JBIC Journal of Biological Inorganic Chemistry

, Volume 10, Issue 6, pp 625–635

Structure and coordination of CuB in the Acidianus ambivalens aa3 quinol oxidase heme–copper center

  • Tiago M. Bandeiras
  • Manuel M. Pereira
  • Miguel Teixeira
  • Pierre Moenne-Loccoz
  • Ninian J. Blackburn
Original Article


The coordination environment of the CuB center of the quinol oxidase from Acidianus ambivalens, a type B heme–copper oxygen reductase, was investigated by Fourier transform (FT) IR and extended X-ray absorption fine structure (EXAFS) spectroscopy. The comparative structural chemistry of dinuclear Fe–Cu sites of the different types of oxygen reductases is of great interest. Fully reduced A. ambivalens quinol oxidase binds CO at the heme a3 center, with ν(CO)=1,973 cm−1. On photolysis, the CO migrated to the CuB center, forming a CuBI–CO complex with ν(CO)=2,047 cm−1. Raising the temperature of the samples to 25°C did not result in a total loss of signal in the FTIR difference spectrum although the intensity of these signals was reduced sevenfold. This observation is consistent with a large energy barrier against the geminate rebinding of CO to the heme iron from CuB, a restricted limited access at the active-site pocket for a second binding, and a kinetically stable CuB–CO complex in A. ambivalens aa3. The CuB center was probed in a number of different states using EXAFS spectroscopy. The oxidized state was best simulated by three histidines and a solvent O scatterer. On reduction, the site became three-coordinate, but in contrast to the bo3 enzyme, there was no evidence for heterogeneity of binding of the coordinated histidines. The CuB centers in both the oxidized and the reduced enzymes also appeared to contain substoichiometric amounts (0.2 mol equiv) of nonlabile chloride ion. EXAFS data of the reduced carbonylated enzyme showed no difference between dark and photolyzed forms. The spectra could be well fit by 2.5 imidazoles, 0.5 Cl and 0.5 CO ligands. This arrangement of scatterers would be consistent with about half the sites remaining as unligated Cu(his)3 and half being converted to Cu(his)2ClCO, a 50/50 ratio of Cu(his)2Cl and Cu(his)3CO, or some combination of these formulations.


Cytochrome c oxidase Heme–copper Quinol oxidase Acidianus ambivalens Extremophile 

Supplementary material

775_2005_12_MOESM1_ESM.pdf (145 kb)
Supplementary material


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Copyright information

© SBIC 2005

Authors and Affiliations

  • Tiago M. Bandeiras
    • 1
  • Manuel M. Pereira
    • 1
  • Miguel Teixeira
    • 1
  • Pierre Moenne-Loccoz
    • 2
  • Ninian J. Blackburn
    • 2
  1. 1.Instituto de Tecnologia Quìmica e BiológicaUniversidade Nova de LisboaOeirasPortugal
  2. 2.Department of Environmental and Biomolecular SystemsOGI School of Science and Engineering at OHSUBeavertonUSA

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