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

Research output: Contribution to journal › Article › peer-review

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). [Abstract copyright: © 2022 The Authors.]

Original language	English
Article number	101312
Journal	STAR protocols
Volume	3
Issue number	2
Early online date	13 Apr 2022
DOIs	https://doi.org/10.1016/j.xpro.2022.101312
Publication status	Published - 17 Jun 2022

Bibliographical note

© 2022 The Authors. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).

Funding Information:
The authors acknowledge the Knut and Alice Wallenberg Foundation (to P.G. through KAW 2015.0131 and 2020.0194); the Lundbeck Foundation (to P.G. through R133-A12689 and R313-2019-774); the Independent Research Fund Denmark (to P.G. through 4183-00559); NordForsk (to P.G. through 82000); the Swedish Research Council (to S.T-H. through 2013-05945 and 2019-06106); the Crafoord Foundation (to S.T.-H. through 20140811 and 20180916); the Magnus Bergvall Foundation (to S.T-H. through 2015-01534); UK Biotechnology & Biosciences Research Council (to R.M.B. and P.K. through BB/P025927/1); P.K. is the recipient of an Aston University 50th Anniversary Prize Fellowship: M.M.S. is supported through funding by Leverhulme Trust UK (ECF-2021-602 ).

S.T.H. P.G. and J.W.M. conceived the study and devised the experimental strategy. J.W.M. and J.H.S. performed the experiments and analyzed the data. All authors contributed to manuscript writing and approved the final version. The authors declare no competing interests.

Keywords

Water - metabolism
Cell Membrane
Single-molecule Assays
Proteolipids - metabolism
Permeability
Aquaporins - metabolism
Protein Biochemistry

Access to Document

10.1016/j.xpro.2022.101312Licence: CC BY 4.0

Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup
© 2022 The Authors. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Final published version, 2.05 MBLicence: CC BY 4.0

Cite this

@article{33d379d8bd7c40f68c23cba5705c0e98,

title = "Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup",

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). [Abstract copyright: {\textcopyright} 2022 The Authors.]",

keywords = "Water - metabolism, Cell Membrane, Single-molecule Assays, Proteolipids - metabolism, Permeability, Aquaporins - metabolism, Protein Biochemistry",

author = "Steffen, {Jonas Hyld} and Missel, {Julie Winkel} and Tamim Al-Jubair and Philip Kitchen and Salman, {Mootaz M} and Bill, {Roslyn M} and Susanna T{\"o}rnroth-Horsefield and Pontus Gourdon",

note = "{\textcopyright} 2022 The Authors. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/). Funding Information: The authors acknowledge the Knut and Alice Wallenberg Foundation (to P.G. through KAW 2015.0131 and 2020.0194); the Lundbeck Foundation (to P.G. through R133-A12689 and R313-2019-774); the Independent Research Fund Denmark (to P.G. through 4183-00559); NordForsk (to P.G. through 82000); the Swedish Research Council (to S.T-H. through 2013-05945 and 2019-06106); the Crafoord Foundation (to S.T.-H. through 20140811 and 20180916); the Magnus Bergvall Foundation (to S.T-H. through 2015-01534); UK Biotechnology & Biosciences Research Council (to R.M.B. and P.K. through BB/P025927/1); P.K. is the recipient of an Aston University 50th Anniversary Prize Fellowship: M.M.S. is supported through funding by Leverhulme Trust UK (ECF-2021-602 ). S.T.H. P.G. and J.W.M. conceived the study and devised the experimental strategy. J.W.M. and J.H.S. performed the experiments and analyzed the data. All authors contributed to manuscript writing and approved the final version. The authors declare no competing interests.",

year = "2022",

month = jun,

day = "17",

doi = "10.1016/j.xpro.2022.101312",

language = "English",

volume = "3",

number = "2",

}

TY - JOUR

T1 - Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup

AU - Steffen, Jonas Hyld

AU - Missel, Julie Winkel

AU - Al-Jubair, Tamim

AU - Kitchen, Philip

AU - Salman, Mootaz M

AU - Bill, Roslyn M

AU - Törnroth-Horsefield, Susanna

AU - Gourdon, Pontus

N1 - © 2022 The Authors. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/). Funding Information: The authors acknowledge the Knut and Alice Wallenberg Foundation (to P.G. through KAW 2015.0131 and 2020.0194); the Lundbeck Foundation (to P.G. through R133-A12689 and R313-2019-774); the Independent Research Fund Denmark (to P.G. through 4183-00559); NordForsk (to P.G. through 82000); the Swedish Research Council (to S.T-H. through 2013-05945 and 2019-06106); the Crafoord Foundation (to S.T.-H. through 20140811 and 20180916); the Magnus Bergvall Foundation (to S.T-H. through 2015-01534); UK Biotechnology & Biosciences Research Council (to R.M.B. and P.K. through BB/P025927/1); P.K. is the recipient of an Aston University 50th Anniversary Prize Fellowship: M.M.S. is supported through funding by Leverhulme Trust UK (ECF-2021-602 ). S.T.H. P.G. and J.W.M. conceived the study and devised the experimental strategy. J.W.M. and J.H.S. performed the experiments and analyzed the data. All authors contributed to manuscript writing and approved the final version. The authors declare no competing interests.

PY - 2022/6/17

Y1 - 2022/6/17

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). [Abstract copyright: © 2022 The Authors.]

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). [Abstract copyright: © 2022 The Authors.]

KW - Water - metabolism

KW - Cell Membrane

KW - Single-molecule Assays

KW - Proteolipids - metabolism

KW - Permeability

KW - Aquaporins - metabolism

KW - Protein Biochemistry

UR - https://www.sciencedirect.com/science/article/pii/S2666166722001927?via%3Dihub

U2 - 10.1016/j.xpro.2022.101312

DO - 10.1016/j.xpro.2022.101312

M3 - Article

C2 - 35496800

SN - 2666-1667

VL - 3

JO - STAR protocols

JF - STAR protocols

IS - 2

M1 - 101312

ER -

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

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

Cite this