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Membrane Protein Production in the Yeast P. pastoris

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Heterologous Expression of Membrane Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2507))

Abstract

The first crystal structures of recombinant mammalian membrane proteins were solved using high-quality protein that had been produced in yeast cells. One of these, the rat Kv1.2 voltage-gated potassium channel, was synthesized in Pichia pastoris. Since then, this yeast species has remained a consistently popular choice of host for synthesizing eukaryotic membrane proteins because it is quick, easy, and cheap to culture and is capable of posttranslational modification. Very recent structures of recombinant membrane proteins produced in P. pastoris include a series of X-ray crystallography structures of the human vitamin K epoxide reductase and a cryo-electron microscopy structure of the TMEM206 proton-activated chloride channel from pufferfish. P. pastoris has also been used to structurally and functionally characterize a range of membrane proteins including tetraspanins, aquaporins, and G protein-coupled receptors. This chapter provides an overview of the methodological approaches underpinning these successes.

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Abbreviations

BMGY:

Buffered glycerol-complex medium

BMMY:

Buffered methanol-complex medium

BSM:

Basal salts medium

DO:

Dissolved oxygen

HPLC:

High-performance liquid chromatography

v/v:

Volume by volume

w/v:

Weight by volume

YPD:

Yeast extract peptone dextrose medium

YPG:

Yeast extract peptone glycerol medium

References

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Acknowledgments

We acknowledge funding from the ERACoBioTech MeMBrane project and BBSRC (BB/R02152X/1) to A.D.G., A.J.R., and R.M.B.

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Authors and Affiliations

  1. Faculty of Health and Life Sciences, Coventry University, Coventry, UK

    Hoor Ayub & Michelle Clare

  2. College of Health and Life Sciences, Aston University, Birmingham, UK

    Luke Broadbent, John Simms, Alan D. Goddard, Alice J. Rothnie & Roslyn M. Bill

Authors
  1. Hoor Ayub
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  2. Michelle Clare
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  3. Luke Broadbent
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  4. John Simms
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  5. Alan D. Goddard
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  6. Alice J. Rothnie
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  7. Roslyn M. Bill
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Corresponding author

Correspondence to Roslyn M. Bill .

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Editors and Affiliations

  1. Université Côte d’Azur, CNRS UMR7275, Institut de Pharmacologie Moléculaire et Cellulaire, Valbonne, France

    Isabelle Mus-Veteau

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Ayub, H. et al. (2022). Membrane Protein Production in the Yeast P. pastoris. In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology, vol 2507. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2368-8_10

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  • DOI: https://doi.org/10.1007/978-1-0716-2368-8_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2367-1

  • Online ISBN: 978-1-0716-2368-8

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