Abstract：
Structural insights into macromolecular and protein complexes provide key clues about the molecular basis of the function. Cryogenic electron microscopy (cryo-EM) has emerged as a powerful structural biology method for studying protein and macromolecular structures at high resolution in both native and near-native states. Despite the ability to get detailed structural insights into the processes underlying protein function using cryo-EM, there has been hesitancy amongst plant biologists to apply the method for biomolecular interaction studies. This is largely evident from the relatively fewer structural depositions of proteins and protein complexes from plant origin in electron microscopy databank. Even though the progress has been slow, cryo-EM has significantly contributed to our understanding of the molecular biology processes underlying photosynthesis, energy transfer in plants, besides viruses infecting plants. This chapter introduces sample preparation for both negative-staining electron microscopy (NSEM) and cryo-EM for plant proteins and macromolecular complexes and data analysis using single particle analysis for beginners.
摘要：
对大分子和蛋白质复合物的结构见解提供了有关功能的分子基础的关键线索。低温电子显微镜（cryo-EM）已成为一种强大的结构生物学方法，可在天然和近天然状态下以高分辨率研究蛋白质和大分子结构。尽管能够使用cryo-EM获得对蛋白质功能基础过程的详细结构见解，植物生物学家对将该方法应用于生物分子相互作用研究一直犹豫不决。从电子显微镜数据库中来自植物来源的蛋白质和蛋白质复合物的相对较少的结构沉积中可以明显看出这一点。尽管进展缓慢,cryo-EM对我们对光合作用的分子生物学过程的理解做出了重大贡献，植物中的能量传递，除了病毒感染植物。本章介绍了用于植物蛋白质和大分子复合物的负染色电子显微镜（NSEM）和cryo-EM的样品制备以及使用初学者的单粒子分析进行数据分析。
