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

Journal: STAR protocols 2022;3(2):101312

PMID: 35496800

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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).

Address: Department of Biomedical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark.; Department of Biochemistry and Structural Biology, Lund University, P.O. Box 124, 221 00 Lund, Sweden.; College of Health and Life Sciences, Aston University, Aston Triangle, B4 7ET Birmingham, UK.; Department of Physiology, Anatomy and Genetics, Kavli Institute for NanoScience Discovery, University of Oxford, Parks Road, OX1 3PT Oxford, UK.; Oxford Parkinson's Disease Centre, University of Oxford, Oxford, UK.; Department of Experimental Medical Science, Lund University, P.O. Box 118, 221 00 Lund, Sweden.

DOI: 10.1016/j.xpro.2022.101312

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MeSH Terms: Aquaporins, Permeability, Proteolipids, Water