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Inducible release of particulates from liposomes using the mechanosensitive channel of large conductance and l-α-lysophosphatidylcholine

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Abstract

The mechanosensitive channel of large conductance (MscL) from Escherichia coli is a prototype for the mechanosensitive class of ion channels and opens one of the largest known gated transmembrane pores. As such, MscL offers the structural framework for the development of liposomal nanovalves for biotechnological applications. Here we incorporated MscL into liposomes and investigated the effects of l-α-lysophosphatidylcholine (LPC) with varying acyl chain lengths or saturation on its pore gating. This was measured by the efflux of encapsulated 5,6-carboxyfluorescein (CF) from the MscL proteoliposomes. Efflux improved in the presence of shorter and double-bonded LPC acyl chains. It was also dependent on the detergent concentration employed during MscL purification. MscL purified in 2 mM dodecyl β-d-maltopyranoside (DDM) had a marked increase in CF efflux compared to MscL purified in 1 mM DDM when treated with LPC. The purification conditions also resulted in increased efflux from proteoliposomes containing the G22C-MscL pore mutant channel, which requires higher membrane tension for its activation compared to WT-MscL.

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Acknowledgments

We thank Dr Charles Cranfield, Dr. Takeshi Nomura, Paul Rohde, and Maryrose Constantine for fruitful discussions and technical help. This study was funded in part by National Health and Medical Research Council of Australia grants to BH and BM and an Australian Postgraduate Award to AF.

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Authors and Affiliations

  1. Institute for Molecular Biosciences, The University of Queensland, St. Lucia, QLD, 4072, Australia

    Alexander Foo, Gamma Chi, Ben Hankamer & Michael J. Landsberg

  2. Menzies Health Institute Queensland and School of Pharmacy, Griffith University, Gold Coast Campus, Queensland, 4222, Australia

    Andrew R. Battle

  3. School of Biomedical Sciences, The University of Queensland, St. Lucia, Queensland, 4072, Australia

    Andrew R. Battle

  4. The Victor Chang Cardiac Research Institute, Lowy Packer Building, 405 Liverpool St, Darlinghurst, NSW, 2010, Australia

    Boris Martinac

  5. St. Vincent’s Clinical School, The University of New South Wales, Darlinghurst, NSW, 2010, Australia

    Boris Martinac

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  1. Alexander Foo
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  2. Andrew R. Battle
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  3. Gamma Chi
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  4. Ben Hankamer
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  5. Michael J. Landsberg
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  6. Boris Martinac
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Correspondence to Boris Martinac.

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Special issue: Biophysics of Mechanotransduction.

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Foo, A., Battle, A.R., Chi, G. et al. Inducible release of particulates from liposomes using the mechanosensitive channel of large conductance and l-α-lysophosphatidylcholine. Eur Biophys J 44, 521–530 (2015). https://doi.org/10.1007/s00249-015-1055-4

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  • DOI: https://doi.org/10.1007/s00249-015-1055-4

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