Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup
(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).
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/c0e959db-ce7c-4905-a3e1-9d3e1260e714
- author
- 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
- organization
- publishing date
- 2022-06-17
- 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
- 2024-04-04 10:17:26
@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}}, }