Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, Bethesda, Maryland 20814, USA; email:
University of British Columbia, Vancouver, British Columbia V6T 1Z2, Canada; email:
Annu Rev Biomed Eng. 2019 Jun 4;21:395-415. doi: 10.1146/annurev-bioeng-060418-052453. Epub 2019 Mar 20.
In recent years, cryo electron microscopy (cryo-EM) technology has been transformed with the development of better instrumentation, direct electron detectors, improved methods for specimen preparation, and improved software for data analysis. Analyses using single-particle cryo-EM methods have enabled determination of structures of proteins with sizes smaller than 100 kDa and resolutions of ∼2 Å in some cases. The use of electron tomography combined with subvolume averaging is beginning to allow the visualization of macromolecular complexes in their native environment in unprecedented detail. As a result of these advances, solutions to many intractable challenges in structural and cell biology, such as analysis of highly dynamic soluble and membrane-embedded protein complexes or partially ordered protein aggregates, are now within reach. Recent reports of structural studies of G protein-coupled receptors, spliceosomes, and fibrillar specimens illustrate the progress that has been made using cryo-EM methods, and are the main focus of this review.
近年来，随着更好的仪器设备、直接电子探测器、改进的样品制备方法以及数据分析改进软件的发展，冷冻电子显微镜（cryo-EM）技术得到了转变。使用单颗粒 cryo-EM 方法的分析已经能够确定小于 100 kDa 的蛋白质的结构，并且在某些情况下分辨率达到 ∼2 Å。电子断层扫描与子体积平均的结合使用开始允许以前所未有的细节可视化在其天然环境中的大分子复合物。由于这些进展，结构和细胞生物学中许多棘手挑战的解决方案，例如分析高度动态的可溶性和膜嵌入的蛋白质复合物或部分有序的蛋白质聚集体，现在已经触手可及。最近关于 G 蛋白偶联受体、剪接体和纤维状标本的结构研究报告说明了使用 cryo-EM 方法取得的进展，这也是本综述的主要重点。
