title: Integral Membrane Protein Expression in Saccharomyces cerevisiae.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA.
authors.forename: Rebba C
authors.initials: RC
authors.lastname: Boswell-Casteel
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA.
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Johnson
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA, 94143, USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Stroud
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA. Franklin-Hays@ouhsc.edu.|Stephenson Oklahoma Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA. Franklin-Hays@ouhsc.edu.
authors.forename: Franklin A
authors.initials: FA
authors.lastname: Hays
mesh_terms: D003001:Cloning, Molecular; D015870:Gene Expression; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development*
chemical_list: D008565:Membrane Proteins; D011994:Recombinant Proteins
keywords: Integral membrane protein; Protein expression; Protein overproduction; Saccharomyces cerevisiae; Yeast