Yeast as a tool for membrane protein production and structure determination

Antonio Carlesso, Raquel Delgado, Oriol Ruiz Isant, Owens Uwangue, Dylan Valli, Roslyn M Bill, Kristina Hedfalk*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Membrane proteins are challenging targets to functionally and structurally characterize. An enduring bottleneck in their study is the reliable production of sufficient yields of stable protein. Here, we evaluate all eukaryotic membrane protein production experiments that have supported the deposition of a high-resolution structure. We focused on the most common yeast host systems, Saccharomyces cerevisiae and Pichia pastoris. The first high-resolution structure of a membrane protein produced in yeast was described in 1999 and today there are 186 structures of α-helical membrane proteins, representing 101 unique proteins from 37 families. Homologous and heterologous production are equally common in S. cerevisiae, while heterologous production dominates in P. pastoris, especially of human proteins, which represent about one-third of the total. Investigating protein engineering approaches (78 proteins from seven families) demonstrated that the majority contained a polyhistidine tag for purification, typically at the C-terminus of the protein. Codon optimization and truncation of hydrophilic extensions were also common approaches to improve yields. We conclude that yeast remains a useful production host for the study of α-helical membrane proteins.

Original languageEnglish
Number of pages14
JournalFEMS Yeast Research
Volume22
Issue number1
DOIs
Publication statusPublished - 20 Oct 2022

Bibliographical note

© The Author(s) 2022. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited.

Keywords

  • Membrane Protein Production
  • Saccharomyces Cerevisiae
  • Pichia Pastoris

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