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/Report/Conference proceedingChapterpeer-review

1 Citation (Scopus)

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. [Abstract copyright: © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.]
Original languageEnglish
Title of host publicationHeterologous Expression of Membrane Proteins. Methods in Molecular Biology
Editors Isabelle Mus-Veteau
PublisherHumana Press
Pages187-199
Number of pages13
Volume2507
ISBN (Electronic)978-1-0716-2368-8
ISBN (Print)978-1-0716-2367-1
DOIs
Publication statusPublished - 1 Jul 2022

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press
ISSN (Print)1064-3745

Keywords

  • Komagataella pastoris
  • Cryoelectron Microscopy
  • Membrane Proteins - metabolism
  • Pichia - genetics - metabolism
  • Rats
  • Recombinant Proteins - chemistry
  • Animals
  • Methanol
  • Yeast
  • AOX1
  • Recombinant membrane protein

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