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High-yield overproduction and purification of human aquaporins from Pichia pastoris

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 (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).

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author
; ; ; ; ; ; and
organization
publishing date
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
2024-06-13 07:16:33
@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 &gt;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}},
}