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Electron and proton transfer in the ba 3 oxidase from Thermus thermophilus

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Abstract

The ba 3-type cytochrome c oxidase from Thermus thermophilus is phylogenetically very distant from the aa 3–type cytochrome c oxidases. Nevertheless, both types of oxidases have the same number of redox-active metal sites and the reduction of O2 to water is catalysed at a haem a 3-CuB catalytic site. The three-dimensional structure of the ba 3 oxidase reveals three possible proton-conducting pathways showing very low homology compared to those of the mitochondrial, Rhodobacter sphaeroides and Paracoccus denitrificans aa 3 oxidases. In this study we investigated the oxidative part of the catalytic cycle of the ba 3 -cytochrome c oxidase using the flow-flash method. After flash-induced dissociation of CO from the fully reduced enzyme in the presence of oxygen we observed rapid oxidation of cytochrome b (k≅6.8 × 104 s−1) and formation of the peroxy (PR) intermediate. In the next step a proton was taken up from solution with a rate constant of ∼1.7 × 104 s−1, associated with formation of the ferryl (F) intermediate, simultaneous with transient reduction of haem b. Finally, the enzyme was oxidized with a rate constant of ∼1,100 s−1, accompanied by additional proton uptake. The total proton uptake stoichiometry in the oxidative part of the catalytic cycle was ~1.5 protons per enzyme molecule. The results support the earlier proposal that the PR and F intermediate spectra are similar (Siletsky et al. Biochim Biophys Acta 1767:138, 2007) and show that even though the architecture of the proton-conducting pathways is different in the ba 3 oxidases, the proton-uptake reactions occur over the same time scales as in the aa 3-type oxidases.

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Authors and Affiliations

  1. A.N.Belozersky Institute of Physicochemical Biology, Moscow State University, Moscow, 119899, Russia

    Irina A. Smirnova & Dmitry Zaslavsky

  2. Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA, 92037, USA

    James A. Fee

  3. Department. of Biochemistry, University of Illinois, Urbana, IL, 61801, USA

    Robert B. Gennis

  4. Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, 106 91, Stockholm, Sweden

    Peter Brzezinski

Authors
  1. Irina A. Smirnova
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  2. Dmitry Zaslavsky
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  3. James A. Fee
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  4. Robert B. Gennis
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  5. Peter Brzezinski
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Correspondence to Peter Brzezinski.

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Smirnova and Zaslavsky contributed equally to the work described in this paper.

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Smirnova, I.A., Zaslavsky, D., Fee, J.A. et al. Electron and proton transfer in the ba 3 oxidase from Thermus thermophilus . J Bioenerg Biomembr 40, 281–287 (2008). https://doi.org/10.1007/s10863-008-9157-7

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  • DOI: https://doi.org/10.1007/s10863-008-9157-7

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