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

Research output: Contribution to journalJournal articleResearchpeer-review

1 Citation (Scopus)
25 Downloads (Pure)

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 languageEnglish
Article number101312
JournalSTAR Protocols
Volume3
Issue number2
Number of pages11
ISSN2666-1667
DOIs
Publication statusPublished - 2022

Bibliographical note

Publisher Copyright:
© 2022 The Authors

Keywords

  • Cell Membrane
  • Protein Biochemistry
  • Single-molecule Assays

Cite this