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The effect of detergents and lipids on the properties of the outer-membrane protein OmcA from Shewanella oneidensis

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Abstract

The protein OmcA from the bacterium Shewanella oneidensis was purified to homogeneity, and characterized using UV–vis spectroelectrochemistry, EPR, and circular dichroism (far- and near-UV regions). EPR spectroscopy showed that many different c-type hemes are present in the protein: high-spin, low-spin, and highly anisotropic low-spin hemes were all detected. The protein was shown to bind tightly to artificial membranes (liposomes) composed of dioleoylphosphatidylglycerol and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (60:40 molar ratio), which mimic the natural environment. However, on the basis of the spectroscopic and electrochemical studies, binding to liposomes does not appear to significantly alter either the structure or the properties of OmcA. On the other hand, the electrochemical properties of OmcA are noticeably changed in the presence of the detergents used during the initial purification stages. In particular, the reduction potentials of two out of the ten OmcA hemes appear to shift in the presence of detergent, perhaps because these hemes are solvent-exposed.

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Acknowledgments

This publication was made possible by support from the National Institutes of Environmental Health Sciences (NIEHS, grant no. 1 R15 ES013955-01). W.E.A. acknowledges National Biomedical EPR Center Grant EB0011980 from NIH.

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Authors and Affiliations

  1. Department of Biophysics, Medical College of Wisconsin, Milwaukee, USA

    Gregory J. Bodemer, William A. Antholine & Liana V. Basova

  2. Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, USA

    Daad Saffarini

  3. Department of Chemistry and Biochemistry, University of Wisconsin-Milwaukee, Milwaukee, USA

    A. Andrew Pacheco

Authors
  1. Gregory J. Bodemer
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  2. William A. Antholine
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  3. Liana V. Basova
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  4. Daad Saffarini
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  5. A. Andrew Pacheco
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Corresponding authors

Correspondence to William A. Antholine or A. Andrew Pacheco.

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Bodemer, G.J., Antholine, W.A., Basova, L.V. et al. The effect of detergents and lipids on the properties of the outer-membrane protein OmcA from Shewanella oneidensis . J Biol Inorg Chem 15, 749–758 (2010). https://doi.org/10.1007/s00775-010-0643-0

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