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

Tamim Al-Jubair; Jonas Hyld Steffen; Julie Winkel Missel; Philip Kitchen; Mootaz M. Salman; Roslyn M. Bill; Pontus Gourdon; Susanna Törnroth-Horsefield

doi:10.1016/j.xpro.2022.101298

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

Tamim Al-Jubair^*, Jonas Hyld Steffen, Julie Winkel Missel, Philip Kitchen, Mootaz M. Salman, Roslyn M. Bill, Pontus Gourdon, Susanna Törnroth-Horsefield

^*Corresponding author for this work

Molecular Cardiology and Membrane Proteins

Research output: Contribution to journal › Journal article › Research › peer-review

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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).

Original language	English
Article number	101298
Journal	STAR Protocols
Volume	3
Issue number	2
Number of pages	19
ISSN	2666-1667
DOIs	https://doi.org/10.1016/j.xpro.2022.101298
Publication status	Published - 2022

Bibliographical note

Publisher Copyright:
© 2022 The Authors

Keywords

Cell Membrane
Molecular Biology
Protein Biochemistry
Protein expression and purification

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High-yield overproduction and purification of human aquaporins from Pichia pastoris. / Al-Jubair, Tamim; Steffen, Jonas Hyld ; Missel, Julie Winkel; Kitchen, Philip; Salman, Mootaz M.; Bill, Roslyn M.; Gourdon, Pontus; Törnroth-Horsefield, Susanna.

In: STAR Protocols, Vol. 3, No. 2, 101298, 2022.

Research output: Contribution to journal › Journal article › Research › peer-review

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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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AU - Gourdon, Pontus

AU - Törnroth-Horsefield, Susanna

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

Abstract

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Keywords

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