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Heterologous Expression of Membrane Proteins pp 23–35Cite as

Membrane Protein Production in the Yeast, S. cerevisiae

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1432))

Abstract

The first crystal structures of recombinant mammalian membrane proteins were solved in 2005 using protein that had been produced in yeast cells. One of these, the rabbit Ca2+-ATPase SERCA1a, was synthesized in Saccharomyces cerevisiae. All host systems have their specific advantages and disadvantages, but yeast has remained a consistently popular choice in the eukaryotic membrane protein field because it is quick, easy and cheap to culture, whilst being able to post-translationally process eukaryotic membrane proteins. Very recent structures of recombinant membrane proteins produced in S. cerevisiae include those of the Arabidopsis thaliana NRT1.1 nitrate transporter and the fungal plant pathogen lipid scramblase, TMEM16. This chapter provides an overview of the methodological approaches underpinning these successes.

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Abbreviations

BCA:

Bicinchoninic acid

BSA:

Bovine serum albumin

CCD:

Charge-coupled device

DoE:

Design of experiments

GFP:

Green fluorescent protein

GOI:

Gene of interest

GPCR:

G protein-coupled receptor

h:

Hour

LioAc:

Lithium acetate

PGAL1 :

GAL1 promoter

PEG:

Polyethylene glycol

s:

Second

T4L:

T4 lysozyme

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Acknowledgments

RMB acknowledges funding from the Biotechnology and Biological Sciences Research Council (BBSRC; via grants BB/I019960/1 and BB/L502194/1) and the Innovative Medicines Joint Undertaking under Grant Agreement number 115583 to the ND4BB ENABLE Consortium.

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Authors and Affiliations

  1. School of Life & Health Sciences, Aston University, Birmingham, B4 7ET, UK

    Stephanie P. Cartwright, Lina Mikaliunaite & Roslyn M. Bill

Authors
  1. Stephanie P. Cartwright
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  2. Lina Mikaliunaite
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  3. 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, IPMC, Valbonne, France

    Isabelle Mus-Veteau

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Cartwright, S.P., Mikaliunaite, L., Bill, R.M. (2016). Membrane Protein Production in the Yeast, S. cerevisiae . In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology, vol 1432. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3637-3_2

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  • DOI: https://doi.org/10.1007/978-1-4939-3637-3_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3635-9

  • Online ISBN: 978-1-4939-3637-3

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