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Patch-Clamp Recordings of the KcsA K+ Channel in Unilamellar Blisters

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

Abstract

Patch-clamp electrophysiology is the standard technique used for the high-resolution functional measurements on ion channels. While studies using patch clamp are commonly carried out following ion channel expression in a heterologous system such as Xenopus oocytes or tissue culture cells, these studies can also be carried out using ion channels reconstituted into lipid vesicles. In this chapter, we describe the methodology for reconstituting ion channels into liposomes and the procedure for the generation of unilamellar blisters from these liposomes that are suitable for patch clamp. Here, we focus on the bacterial K+ channel KcsA, although the methodologies described in this chapter should be applicable for the functional analysis of other ion channels.

Key words

  • GUV recordings
  • KcsA
  • Lipid vesicles
  • Blisters
  • Patch clamp

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Acknowledgment

This research was supported by a grant from the NIH (GM087546) to F.I.V.

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Correspondence to Francis I. Valiyaveetil .

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Matulef, K., Valiyaveetil, F.I. (2018). Patch-Clamp Recordings of the KcsA K+ Channel in Unilamellar Blisters. 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_14

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  • DOI: https://doi.org/10.1007/978-1-4939-7362-0_14

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