Literature DB >> 27485336

Integral Membrane Protein Expression in Saccharomyces cerevisiae.

Rebba C Boswell-Casteel1, Jennifer M Johnson1, Robert M Stroud2, Franklin A Hays3,4.   

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.

Entities:  

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

Mesh:

Substances:

Year:  2016        PMID: 27485336      PMCID: PMC6166409          DOI: 10.1007/978-1-4939-3637-3_11

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  12 in total

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Authors:  V L Price; W E Taylor; W Clevenger; M Worthington; E T Young
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4.  Evaluation of the Saccharomyces cerevisiae ADH2 promoter for protein synthesis.

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Journal:  Yeast       Date:  2005-04-30       Impact factor: 3.239

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7.  Overexpression and purification of integral membrane proteins in yeast.

Authors:  Franklin A Hays; Zygy Roe-Zurz; Robert M Stroud
Journal:  Methods Enzymol       Date:  2010-03-01       Impact factor: 1.600

8.  GFP-based optimization scheme for the overexpression and purification of eukaryotic membrane proteins in Saccharomyces cerevisiae.

Authors:  David Drew; Simon Newstead; Yo Sonoda; Hyun Kim; Gunnar von Heijne; So Iwata
Journal:  Nat Protoc       Date:  2008       Impact factor: 13.491

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Journal:  Proc Natl Acad Sci U S A       Date:  2007-08-20       Impact factor: 11.205

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Authors:  Min Li; Franklin A Hays; Zygy Roe-Zurz; Linda Vuong; Libusha Kelly; Chi-Min Ho; Renée M Robbins; Ursula Pieper; Joseph D O'Connell; Larry J W Miercke; Kathleen M Giacomini; Andrej Sali; Robert M Stroud
Journal:  J Mol Biol       Date:  2008-11-24       Impact factor: 5.469

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  2 in total

1.  Overproduction of Membrane-Associated, and Integrated, Proteins Using Saccharomyces cerevisiae.

Authors:  Landon Haslem; Marina Brown; Xin A Zhang; Jennifer M Hays; Franklin A Hays
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2.  First purified recombinant CYP75B including transmembrane helix with unexpected high substrate specificity to (2R)-naringenin.

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  2 in total

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