High-yield overproduction and purification of human aquaporins from Pichia pastoris
(2022) In STAR Protocols 3(2).- Abstract
Aquaporins (AQPs) are membrane-bound water channels that play crucial roles in maintaining the water homeostasis of the human body. Here, we present a protocol for high-yield recombinant expression of human AQPs in the methylotropic yeast Pichia pastoris and subsequent AQP purification. The protocol typically yields 1–5 mg AQP per g of yeast cell at >95% purity and is compatible with any membrane protein cloned into Pichia pastoris, although expression levels may vary. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Frick et al. (2014).
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/ae0c4ae0-69f1-4806-91da-463831e7937a
- author
- Al-Jubair, Tamim LU ; Steffen, Jonas Hyld ; Missel, Julie Winkel ; Kitchen, Philip ; Salman, Mootaz M. ; Bill, Roslyn M. ; Gourdon, Pontus LU and Törnroth-Horsefield, Susanna LU
- organization
- publishing date
- 2022
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Cell Membrane, Molecular Biology, Protein Biochemistry, Protein expression and purification
- in
- STAR Protocols
- volume
- 3
- issue
- 2
- article number
- 101298
- publisher
- Cell Press
- external identifiers
-
- scopus:85127974289
- pmid:35463467
- ISSN
- 2666-1667
- DOI
- 10.1016/j.xpro.2022.101298
- language
- English
- LU publication?
- yes
- id
- ae0c4ae0-69f1-4806-91da-463831e7937a
- date added to LUP
- 2022-06-14 11:53:52
- date last changed
- 2024-09-19 16:34:29
@article{ae0c4ae0-69f1-4806-91da-463831e7937a, abstract = {{<p>Aquaporins (AQPs) are membrane-bound water channels that play crucial roles in maintaining the water homeostasis of the human body. Here, we present a protocol for high-yield recombinant expression of human AQPs in the methylotropic yeast Pichia pastoris and subsequent AQP purification. The protocol typically yields 1–5 mg AQP per g of yeast cell at >95% purity and is compatible with any membrane protein cloned into Pichia pastoris, although expression levels may vary. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020) and Frick et al. (2014).</p>}}, author = {{Al-Jubair, Tamim and Steffen, Jonas Hyld and Missel, Julie Winkel and Kitchen, Philip and Salman, Mootaz M. and Bill, Roslyn M. and Gourdon, Pontus and Törnroth-Horsefield, Susanna}}, issn = {{2666-1667}}, keywords = {{Cell Membrane; Molecular Biology; Protein Biochemistry; Protein expression and purification}}, language = {{eng}}, number = {{2}}, publisher = {{Cell Press}}, series = {{STAR Protocols}}, title = {{High-yield overproduction and purification of human aquaporins from Pichia pastoris}}, url = {{http://dx.doi.org/10.1016/j.xpro.2022.101298}}, doi = {{10.1016/j.xpro.2022.101298}}, volume = {{3}}, year = {{2022}}, }