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

doi:10.1016/j.xpro.2022.101312

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

^*Corresponding author for this work

Molecular Cardiology and Membrane Proteins

Research output: Contribution to journal › Journal article › Research › peer-review

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Abstract

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 language	English
Article number	101312
Journal	STAR Protocols
Volume	3
Issue number	2
Number of pages	11
ISSN	2666-1667
DOIs	https://doi.org/10.1016/j.xpro.2022.101312
Publication status	Published - 2022

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Publisher Copyright:
© 2022 The Authors

Keywords

Cell Membrane
Protein Biochemistry
Single-molecule Assays

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Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup. / Steffen, Jonas Hyld ; Missel, Julie Winkel; Al-Jubair, Tamim; Kitchen, Philip; Salman, Mootaz M.; Bill, Roslyn M.; Törnroth-Horsefield, Susanna; Gourdon, Pontus.

In: STAR Protocols, Vol. 3, No. 2, 101312, 2022.

Research output: Contribution to journal › Journal article › Research › peer-review

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abstract = "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).",

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AU - Törnroth-Horsefield, Susanna

AU - Gourdon, Pontus

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N2 - 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).

AB - 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).

KW - Cell Membrane

KW - Protein Biochemistry

KW - Single-molecule Assays

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JO - STAR Protocols

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

Abstract

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Keywords

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