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Modulating bilayer mechanical properties to promote the coupled folding and insertion of an integral membrane protein

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Abstract

Bilayer mechanical properties are not only of crucial importance to the mechanism of action of mechanosensation in lipid membranes but also affect preparative laboratory tasks such as membrane-protein refolding. We report this for coupled refolding and bilayer insertion of outer membrane phospholipase A (OmpLA), an integral membrane enzyme that catalyses the hydrolytic cleavage of glycerophospholipids. OmpLA can be refolded into a variety of detergent micelles and unilamellar vesicles composed of short-chain phospholipids but, in the absence of chemical or molecular chaperones, not into thicker membranes. Controlled modulation of bilayer mechanical properties by judicious use of subsolubilising concentrations of detergents induces monolayer curvature strain, acyl chain fluidisation, membrane thinning, and transient aqueous bilayer defects. This enables quantitative and functional refolding of OmpLA even into bilayer membranes composed of long-chain phospholipids to yield enzymatically active proteoliposomes without requiring membrane solubilisation.

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Acknowledgments

We thank Dr. Carolyn Vargas (University of Kaiserslautern) for helpful comments on the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft (DFG) through International Research Training Group IRTG 1830 and by the Research Initiative BioComp.

Conflict of interest

The authors declare no competing financial interest.

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Authors and Affiliations

  1. Molecular Biophysics, University of Kaiserslautern, Erwin-Schrödinger-Str. 13, 67663, Kaiserslautern, Germany

    Michaela Herrmann, Bartholomäus Danielczak, Martin Textor, Jessica Klement & Sandro Keller

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  1. Michaela Herrmann
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  2. Bartholomäus Danielczak
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  3. Martin Textor
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  4. Jessica Klement
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  5. Sandro Keller
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Correspondence to Sandro Keller.

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Special issue: Biophysics of Mechanotransduction.

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Herrmann, M., Danielczak, B., Textor, M. et al. Modulating bilayer mechanical properties to promote the coupled folding and insertion of an integral membrane protein. Eur Biophys J 44, 503–512 (2015). https://doi.org/10.1007/s00249-015-1032-y

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  • DOI: https://doi.org/10.1007/s00249-015-1032-y

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