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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 LU ; Kitchen, Philip ; Salman, Mootaz M. ; Bill, Roslyn M. ; Törnroth-Horsefield, Susanna LU and Gourdon, Pontus LU (2022) In STAR Protocols 3(2).
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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author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cell Membrane, Protein Biochemistry, Single-molecule Assays
in
STAR Protocols
volume
3
issue
2
article number
101312
publisher
Cell Press
external identifiers
  • pmid:35496800
  • scopus:85128333189
ISSN
2666-1667
DOI
10.1016/j.xpro.2022.101312
language
English
LU publication?
yes
id
c0e959db-ce7c-4905-a3e1-9d3e1260e714
date added to LUP
2022-06-17 15:40:10
date last changed
2024-04-18 12:30:33
@article{c0e959db-ce7c-4905-a3e1-9d3e1260e714,
  abstract     = {{<p>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).</p>}},
  author       = {{Steffen, Jonas Hyld and Missel, Julie Winkel and Al-Jubair, Tamim and Kitchen, Philip and Salman, Mootaz M. and Bill, Roslyn M. and Törnroth-Horsefield, Susanna and Gourdon, Pontus}},
  issn         = {{2666-1667}},
  keywords     = {{Cell Membrane; Protein Biochemistry; Single-molecule Assays}},
  language     = {{eng}},
  month        = {{06}},
  number       = {{2}},
  publisher    = {{Cell Press}},
  series       = {{STAR Protocols}},
  title        = {{Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup}},
  url          = {{http://dx.doi.org/10.1016/j.xpro.2022.101312}},
  doi          = {{10.1016/j.xpro.2022.101312}},
  volume       = {{3}},
  year         = {{2022}},
}