Abstract
Eukaryotic membrane proteins, many of which are key players in various biological processes, constitute more than half of the drug targets and represent important candidates for structural studies. In contrast to their physiological significance, only very limited number of eukaryotic membrane protein structures have been obtained due to the technical challenges in the generation of recombinant proteins. In this review, we examine the major recombinant expression systems for eukaryotic membrane proteins and compare their relative advantages and disadvantages. We also attempted to summarize the recent technical strategies in the advancement of eukaryotic membrane protein purification and crystallization.
Highlights
It is estimated that approximately 30% of the protein-coding genes are for integral membrane proteins (IMPs) in human (Overington et al, 2006; Murray et al, 2012)
IMPs constitute more than 50% of the US Food and Drug Administration (FDA)approved drug targets (Russell and Eggleston, 2000; Yildirim et al, 2007)
In the hope of extracting some general principles on the expression and crystallization of eukaryotic membrane proteins, we examine the expression systems for the eukaryotic IMPs whose structures are obtained, attempt to summarize and compare the advantages and disadvantages of the representative recombinant expression systems, and delineate the detailed information in eukaryotic membrane protein purification and crystallization (Table 1)
Summary
It is estimated that approximately 30% of the protein-coding genes are for integral membrane proteins (IMPs) in human (Overington et al, 2006; Murray et al, 2012). Whereas Escherichia coli proved to be the best host for most of prokaryotic IMPs of known structures, eukaryotic IMPs, with very few exceptions, requires eukaryotic expression systems including yeast, baculovirus-infected insect cells, and mammalian cells (Bill et al, 2011; Snider and Stephen, 2014). The recombinantly expressed eukaryotic IMPs of known structures were obtained from four systems: E. coli, yeasts (Pichia Pastoris and Saccharomyces cerevisiae), insect cells, and mammalian cells. These expression systems have their respective advantages and disadvantages. E. coli C43 (DE3) and C41 (DE3) strains were developed for over-expression of membrane proteins (Miroux and Walker, 1996; Dumon-Seignovert et al, 2004) These E. coli strains were employed to over-express the large majority of prokaryotic IMPs whose structures were obtained.
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