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Transport of Na+ and K+ by an antiporter-related subunit from the Escherichia coli NADH dehydrogenase I produced in Saccharomyces cerevisiae

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Abstract

The NADH dehydrogenase I from Escherichia coli is a bacterial homolog of the mitochondrial complex I which translocates Na+ rather than H+. To elucidate the mechanism of Na+ transport, the C-terminally truncated NuoL subunit (NuoLN) which is related to Na+/H+ antiporters was expressed as a protein A fusion protein (ProtA–NuoLN) in the yeast Saccharomyces cerevisiae which lacks an endogenous complex I. The fusion protein inserted into membranes from the endoplasmatic reticulum (ER), as confirmed by differential centrifugation and Western analysis. Membrane vesicles containing ProtA–NuoLN catalyzed the uptake of Na+ and K+ at rates which were significantly higher than uptake by the control vesicles under identical conditions, demonstrating that ProtA–NuoLN translocated Na+ and K+ independently from other complex I subunits. Na+ transport by ProtA–NuoLN was inhibited by EIPA (5-(N-ethyl-N-isopropyl)-amiloride) which specifically reacts with Na+/H+ antiporters. The cation selectivity and function of the NuoL subunit as a transporter module of the NADH dehydrogenase complex is discussed.

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Abbreviations

EIPA:

5-(N-ethyl-N-isopropyl)-amiloride

TPP+ :

Tetraphenylphosphonium

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Acknowledgments

This work was supported by grants from the Swiss National Science Foundation, the Vontobel Stiftung and Parkinson Schweiz to J.S. C.S. was supported by a postdoctoral fellowship from the Ernst Schering Research Foundation. We thank Roland Lill, University of Marburg, for providing us with the anti-neurospora porin antibody and Markus Aebi, ETH Zürich, for the generous gift of the anti-WBP1 antibody. We also thank Cecilia Hägerhäll, Lund University, for in-depth discussions.

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Authors and Affiliations

  1. Department of Biochemistry, University of Zürich, Winterthurerstrasse 190, 8057, Zürich, Switzerland

    Anja C. Gemperli & Julia Steuber

  2. Institute of Molecular Biology and Biophysics, Eidgenössische Technische Hochschule (ETH), Schafmattstasse 20, 8093, Zürich, Switzerland

    Christiane Schaffitzel

  3. Institute of Microbiology, Eidgenössische Technische Hochschule (ETH), Wolfgang-Pauli-Strasse 10, 8093, Zürich, Switzerland

    Claude Jakob

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  1. Anja C. Gemperli
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  2. Christiane Schaffitzel
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  3. Claude Jakob
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  4. Julia Steuber
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Correspondence to Julia Steuber.

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Communicated by A. Brakhage.

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Gemperli, A.C., Schaffitzel, C., Jakob, C. et al. Transport of Na+ and K+ by an antiporter-related subunit from the Escherichia coli NADH dehydrogenase I produced in Saccharomyces cerevisiae . Arch Microbiol 188, 509–521 (2007). https://doi.org/10.1007/s00203-007-0272-3

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