title: Expression strategies for structural studies of eukaryotic membrane proteins.
abstract: Integral membrane proteins in eukaryotes are central to various cellular processes and key targets in structural biology, biotechnology and drug development. However, the number of available structures for eukaryotic membrane protein belies their physiological importance. Recently, the number of available eukaryotic membrane protein structures has been steadily increasing due to the development of novel strategies in construct design, expression and structure determination. Here, we examine the major expression systems exploited for eukaryotic membrane proteins. Additionally we strive to tabulate and describe the recent expression strategies in eukaryotic membrane protein structural biology. We find that a majority of targets have been expressed in advanced host systems and modified from their wild-type form with distinct focus on conformation and thermostabilisation. However, strategies for native protein purification should also be considered where possible, particularly in light of the recent advances in single particle cryo electron microscopy.
authors.affiliation: Danish Research Institute of Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Aarhus University, Aarhus, Denmark; Centre for Membrane Pumps in Cells and Disease-PUMPkin, Danish National Research Foundation, Aarhus, Denmark; Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Lyons
authors.affiliation: Danish Research Institute of Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Aarhus University, Aarhus, Denmark; Centre for Membrane Pumps in Cells and Disease-PUMPkin, Danish National Research Foundation, Aarhus, Denmark; Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark; EMBL, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Shahsavar
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
authors.forename: Peter Aasted
authors.initials: PA
authors.lastname: Paulsen
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark; Aarhus Institute of Advanced Studies, Aarhus University, Denmark.
authors.forename: Bjørn Panyella
authors.initials: BP
authors.lastname: Pedersen
authors.affiliation: Danish Research Institute of Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Aarhus University, Aarhus, Denmark; Centre for Membrane Pumps in Cells and Disease-PUMPkin, Danish National Research Foundation, Aarhus, Denmark; Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark. Electronic address: pn@mbg.au.dk.
authors.forename: Poul
authors.initials: P
authors.lastname: Nissen
mesh_terms: D000818:Animals; D003460:Crystallization; D003902:Detergents / Q000494:pharmacology; D056890:Eukaryota / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics*
chemical_list: D003902:Detergents; D008565:Membrane Proteins