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Studying Mechanosensitivity of Two-Pore Domain K+ Channels in Cellular and Reconstituted Proteoliposome Membranes

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Potassium Channels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1684))

Abstract

Mechanical force sensation is fundamental to a wide breadth of biology from the classic senses of touch, pain, hearing, and balance to less conspicuous sensations of proprioception, blood pressure, and osmolarity and basic aspects of cell growth, differentiation, and development. These diverse and essential systems use force-gated (or mechanosensitive) ion channels that convert mechanical stimuli into cellular electrical signals. TRAAK, TREK1, and TREK2 are K+-selective ion channels of the two-pore domain K+ (K2P) family that are mechanosensitive: they are gated open by increasing membrane tension. TRAAK and TREK channels are thought to play roles in somatosensory and other mechanosensory processes in neuronal and non-neuronal tissues. Here, we present protocols for three assays to study mechanical activation of these channels in cell membranes: (1) cell swelling, (2) cell poking, and (3) patched membrane stretching. Patched membrane stretching is also applicable to the study of mechanosensitive K2P channel activity in a cell-free system and a procedure for proteoliposome reconstitution and patching is also presented. These approaches are also readily applicable to the study of other mechanosensitive ion channels.

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Author notes

  1. Josefina del Mármol and Robert A. Rietmeijer contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of California – Berkeley, 289 Life Science Addition, Berkeley, CA, 94720, USA

    Josefina del Mármol & Stephen G. Brohawn

  2. The Helen Wills Neuroscience Institute, University of California – Berkeley, 289 Life Science Addition, Berkeley, CA, 94720, USA

    Josefina del Mármol & Stephen G. Brohawn

  3. Biophysics Graduate Group, University of California, Berkeley, CA, 94720, USA

    Robert A. Rietmeijer

Authors
  1. Josefina del Mármol
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  2. Robert A. Rietmeijer
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  3. Stephen G. Brohawn
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Corresponding author

Correspondence to Stephen G. Brohawn .

Editor information

Editors and Affiliations

  1. Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, Oregon, USA

    Show-Ling Shyng

  2. Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, Oregon, USA

    Francis I. Valiyaveetil

  3. The Vollum Institute, Oregon Health and Science University, Portland, Oregon, USA

    Matt Whorton

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del Mármol, J., Rietmeijer, R.A., Brohawn, S.G. (2018). Studying Mechanosensitivity of Two-Pore Domain K+ Channels in Cellular and Reconstituted Proteoliposome Membranes. In: Shyng, SL., Valiyaveetil, F., Whorton, M. (eds) Potassium Channels. Methods in Molecular Biology, vol 1684. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7362-0_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7362-0_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7361-3

  • Online ISBN: 978-1-4939-7362-0

  • eBook Packages: Springer Protocols

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