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Functional production of the Na+ F1FO ATP synthase from Acetobacterium woodii in Escherichia coli requires the native AtpI

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Abstract

The Na+ F1FO ATP synthase of the anaerobic, acetogenic bacterium Acetobacterium woodii has a unique FOVO hybrid rotor that contains nine copies of a FO-like c subunit and one copy of a VO-like c 1 subunit with one ion binding site in four transmembrane helices whose cellular function is obscure. Since a genetic system to address the role of different c subunits is not available for this bacterium, we aimed at a heterologous expression system. Therefore, we cloned and expressed its Na+ F1FO ATP synthase operon in Escherichia coli. A Δatp mutant of E. coli produced a functional, membrane-bound Na+ F1FO ATP synthase that was purified in a single step after inserting a His6-tag to its β subunit. The purified enzyme was competent in Na+ transport and contained the FOVO hybrid rotor in the same stoichiometry as in A. woodii. Deletion of the atpI gene from the A. woodii operon resulted in a loss of the c ring and a mis-assembled Na+ F1FO ATP synthase. AtpI from E. coli could not substitute AtpI from A. woodii. These data demonstrate for the first time a functional production of a FOVO hybrid rotor in E. coli and revealed that the native AtpI is required for assembly of the hybrid rotor.

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Authors and Affiliations

  1. Molecular Microbiology & Bioenergetics, Institute of Molecular Biosciences, Johann Wolfgang Goethe University Frankfurt/Main, Max-von-Laue-Str. 9, 60438, Frankfurt, Germany

    Karsten Brandt, Daniel B. Müller & Volker Müller

  2. Institute for Physical and Theoretical Chemistry, Johann Wolfgang Goethe University Frankfurt/Main, Max-von-Laue Str. 7, 60438, Frankfurt, Germany

    Jan Hoffmann & Bernd Brutschy

  3. Microbiology, Department of Biology/Chemistry, University of Osnabrück, Barbarastraße 11, 49069, Osnabrück, Germany

    Christine Hübert & Gabriele Deckers-Hebestreit

Authors
  1. Karsten Brandt
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  2. Daniel B. Müller
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  3. Jan Hoffmann
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  4. Christine Hübert
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  5. Bernd Brutschy
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  6. Gabriele Deckers-Hebestreit
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  7. Volker Müller
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Correspondence to Volker Müller.

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Karsten Brandt and Daniel B. Müller are authors that contributed equally to this article.

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Brandt, K., Müller, D.B., Hoffmann, J. et al. Functional production of the Na+ F1FO ATP synthase from Acetobacterium woodii in Escherichia coli requires the native AtpI. J Bioenerg Biomembr 45, 15–23 (2013). https://doi.org/10.1007/s10863-012-9474-8

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  • DOI: https://doi.org/10.1007/s10863-012-9474-8

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