Format

Send to

Choose Destination
Methods Mol Biol. 2016;1432:163-86. doi: 10.1007/978-1-4939-3637-3_11.

Integral Membrane Protein Expression in Saccharomyces cerevisiae.

Author information

1
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA.
2
Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA, 94143, USA.
3
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA. Franklin-Hays@ouhsc.edu.
4
Stephenson Oklahoma Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA. Franklin-Hays@ouhsc.edu.

Abstract

Eukaryotic integral membrane proteins are challenging targets for crystallography or functional characterization in a purified state. Since expression is often a limiting factor when studying this difficult class of biological macromolecules, the intent of this chapter is to focus on the expression of eukaryotic integral membrane proteins (IMPs) using the model organism Saccharomyces cerevisiae. S. cerevisiae is a prime candidate for the expression of eukaryotic IMPs because it offers the convenience of using episomal expression plasmids, selection of positive transformants, posttranslational modifications, and it can properly fold and target IMPs. Here we present a generalized protocol and insights based on our collective knowledge as an aid to overcoming the challenges faced when expressing eukaryotic IMPs in S. cerevisiae.

KEYWORDS:

Integral membrane protein; Protein expression; Protein overproduction; Saccharomyces cerevisiae; Yeast

PMID:
27485336
PMCID:
PMC6166409
DOI:
10.1007/978-1-4939-3637-3_11
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Springer Icon for PubMed Central
Loading ...
Support Center