Characterization of human aquaporin protein-protein interactions using microscale thermophoresis (MST)
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Aquaporin water channels (AQPs) are membrane proteins that maintain cellular water homeostasis. The interactions between human AQPs and other proteins play crucial roles in AQP regulation by both gating and trafficking. Here, we describe a protocol for characterizing the interaction between a human AQP and a soluble interaction partner using microscale thermophoresis (MST). MST has the advantage of low sample consumption and high detergent compatibility enabling AQP protein-protein interaction investigation with a high level of control of components and environment. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Roche et al. (2017).
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 101316 |
| Tidsskrift | STAR Protocols |
| Vol/bind | 3 |
| Udgave nummer | 2 |
| Antal sider | 8 |
| ISSN | 2666-1667 |
| DOI | |
| Status | Udgivet - 2022 |
Bibliografisk note
Funding Information:
We acknowledge 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); 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); 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 through funding by Leverhulme Trust (ECF-2021-602 ). S.T.H. and T.A.J. conceived the study and devised the experimental strategy. T.A.J. and S.T.H. performed the experiments and analyzed the data. All authors contributed to manuscript writing and approved the final version. The authors declare no competing interests.
Funding Information:
We acknowledge 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 ); 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 ); 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 through funding by Leverhulme Trust (ECF-2021-602 ).
Publisher Copyright:
© 2022 The Authors
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