Zorman Sarah, Botte Mathieu, Jiang Qiyang, Collinson Ian, Schaffitzel Christiane
European Molecular Biology Laboratory, Grenoble Outstation, Grenoble 38042, France; Unit for Virus Host-Cell Interactions, University Grenoble Alpes-EMBL-CNRS, Grenoble 38042 France.
School of Biochemistry, University of Bristol, Bristol BS8 1TD, United Kingdom.
Curr Opin Struct Biol. 2015 Jun;32:123-30. doi: 10.1016/j.sbi.2015.03.010. Epub 2015 Apr 15.
Most membrane-proteins exist in complexes rather than isolated entities. To fully understand their biological function it is essential to study the intact membrane-protein assemblies. The overexpression and purification of many essential membrane-protein complexes is still a considerable and often unsurmountable challenge. In these cases, extraction from source is the only option for many large multi-subunit cellular machines. Here, we describe recent advances in overexpression of multi-subunit membrane-protein complexes, the strategies to stabilize these complexes and highlight major achievements in membrane-protein structural research that were facilitated by the prospect of achieving subnanometer to near-atomic resolution by electron cryo-microscopy.
大多数膜蛋白以复合物形式存在,而非孤立的个体。要全面理解其生物学功能,研究完整的膜蛋白组装体至关重要。许多重要膜蛋白复合物的过表达和纯化仍是一项艰巨且往往难以克服的挑战。在这些情况下,从来源中提取是许多大型多亚基细胞机器的唯一选择。在此,我们描述了多亚基膜蛋白复合物过表达的最新进展、稳定这些复合物的策略,并强调了膜蛋白结构研究的主要成就,这些成就得益于通过冷冻电子显微镜实现亚纳米至近原子分辨率的前景。