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

Jonas Hyld Steffen, Julie Winkel Missel, Tamim Al-Jubair, Philip Kitchen, Mootaz M Salman, Roslyn M Bill, Susanna Törnroth-Horsefield, Pontus Gourdon

STAR protocols 2022 Jun 17

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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). © 2022 The Authors.

Citation

Jonas Hyld Steffen, Julie Winkel Missel, Tamim Al-Jubair, Philip Kitchen, Mootaz M Salman, Roslyn M Bill, Susanna Törnroth-Horsefield, Pontus Gourdon. Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup. STAR protocols. 2022 Jun 17;3(2):101312