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 |
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Article number | 101298 |
Journal | STAR Protocols |
Volume | 3 |
Issue number | 2 |
Number of pages | 19 |
ISSN | 2666-1667 |
DOIs | |
Publication status | Published - 2022 |
Bibliographical note
Publisher Copyright:© 2022 The Authors
Keywords
- Cell Membrane
- Molecular Biology
- Protein Biochemistry
- Protein expression and purification