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.
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Acknowledgement
This work was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences (NIGMS) of the National Institutes of Health under grant number P20GM103639, Oklahoma Center for the Advancement of Science grant HR11-046 (to F.A.H.), American Heart Association predoctoral fellowship 13PRE17040024 (to R.C.B-C.), and NIGMS grant GM24485 (to RMS).
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Boswell-Casteel, R.C., Johnson, J.M., Stroud, R.M., Hays, F.A. (2016). Integral Membrane Protein Expression in Saccharomyces cerevisiae . In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology, vol 1432. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3637-3_11
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DOI: https://doi.org/10.1007/978-1-4939-3637-3_11
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