Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup

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Aquaporins (AQPs) are water channels embedded in the cell membrane that are critical in maintaining water homeostasis. We describe a protocol for determining the water permeation capacity of AQPs reconstituted into proteoliposomes. Using a stopped-flow setup, AQP embedded in proteoliposomes are exposed to an osmogenic gradient that triggers water flux. The consequent effects on proteoliposome size can be tracked using the fluorescence of an internalized fluorophore. This enables controlled characterization of water flux by AQPs. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020).

Original languageEnglish
Article number101312
JournalSTAR Protocols
Issue number2
Number of pages11
Publication statusPublished - 2022

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© 2022 The Authors

    Research areas

  • Cell Membrane, Protein Biochemistry, Single-molecule Assays

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