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.
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.
Curr Opin Struct Biol. 2016 Jun;38:137-44. doi: 10.1016/j.sbi.2016.06.011. Epub 2016 Jun 27.
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.
真核生物的整合膜蛋白是各种细胞过程的核心，也是结构生物学、生物技术和药物开发的关键靶点。然而，可用的真核膜蛋白结构数量与其生理重要性不成比例。最近，由于构建设计、表达和结构测定的新策略的发展，可用的真核膜蛋白结构数量稳步增加。在这里，我们检查了用于真核膜蛋白的主要表达系统。此外，我们还努力对真核膜蛋白结构生物学中的最新表达策略进行列表和描述。我们发现，大多数靶标都在先进的宿主系统中表达，并从其野生型形式进行了修饰，重点是构象和热稳定性。然而，也应该考虑到天然蛋白质的纯化策略，特别是在单颗粒冷冻电子显微镜技术的最新进展的情况下。
