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

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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 languageEnglish
Article number101298
JournalSTAR Protocols
Issue number2
Number of pages19
Publication statusPublished - 2022

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© 2022 The Authors

    Research areas

  • Cell Membrane, Molecular Biology, Protein Biochemistry, Protein expression and purification

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