Membrane Protein Production in the Yeast P. pastoris

Hoor Ayub; Michelle Clare; Luke Broadbent; John Simms; Alan D Goddard; Alice J Rothnie; Roslyn M Bill

doi:10.1007/978-1-0716-2368-8_10

Membrane Protein Production in the Yeast P. pastoris

Hoor Ayub, Michelle Clare, Luke Broadbent, John Simms, Alan D Goddard, Alice J Rothnie, Roslyn M Bill

Research output: Chapter in Book/Published conference output › Chapter (peer-reviewed) › peer-review

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.

Original language	English
Title of host publication	Methods in Molecular Biology
Editors	I. Mus-Veteau
Publisher	Springer
Pages	187-199
Number of pages	13
Volume	2507
ISBN (Electronic)	978-1-0716-2368-8
ISBN (Print)	978-1-0716-2367-1
DOIs	https://doi.org/10.1007/978-1-0716-2368-8_10
Publication status	Published - 1 Jul 2022

Publication series

Name	Methods in Molecular Biology
Volume	2507
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Bibliographical note

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

Keywords

AOX1
Komagataella pastoris
Methanol
Recombinant membrane protein
Yeast

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10.1007/978-1-0716-2368-8_10

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AU - Bill, Roslyn M

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AB - 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.

KW - AOX1

KW - Komagataella pastoris

KW - Methanol

KW - Recombinant membrane protein

KW - Yeast

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Membrane Protein Production in the Yeast P. pastoris

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