DiMaio F, Chiu W
University of Washington, Seattle, WA, United States; Institute for Protein Design, University of Washington, Seattle, WA, United States.
National Center for Macromolecular Imaging, Baylor College of Medicine, Houston, TX, United States.
Methods Enzymol. 2016;579:255-76. doi: 10.1016/bs.mie.2016.06.003. Epub 2016 Aug 12.
Electron cryo-microscopy (cryoEM) has advanced dramatically to become a viable tool for high-resolution structural biology research. The ultimate outcome of a cryoEM study is an atomic model of a macromolecule or its complex with interacting partners. This chapter describes a variety of algorithms and software to build a de novo model based on the cryoEM 3D density map, to optimize the model with the best stereochemistry restraints and finally to validate the model with proper protocols. The full process of atomic structure determination from a cryoEM map is described. The tools outlined in this chapter should prove extremely valuable in revealing atomic interactions guided by cryoEM data.
电子冷冻显微镜技术(cryoEM)已取得了巨大进展,成为高分辨率结构生物学研究的一种可行工具。cryoEM研究的最终成果是一个大分子或其与相互作用伙伴的复合物的原子模型。本章介绍了多种算法和软件,用于基于cryoEM三维密度图构建从头模型,利用最佳立体化学约束优化模型,并最终通过适当的协议验证模型。描述了从cryoEM图确定原子结构的完整过程。本章概述的工具在揭示由cryoEM数据指导的原子相互作用方面应具有极高的价值。