Rubach Pawel, Majorek Karolina A, Gucwa Michal, Murzyn Krzysztof, Wlodawer Alexander, Minor Wladek
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, USA.
Institute of Information Systems and Digital Economy, Warsaw School of Economics, Warsaw, Poland.
Expert Opin Drug Discov. 2025 Feb;20(2):163-176. doi: 10.1080/17460441.2025.2450636. Epub 2025 Jan 16.
Macromolecular X-ray crystallography (XRC), nuclear magnetic resonance (NMR), and cryo-electron microscopy (cryoEM) are the primary techniques for determining atomic-level, three-dimensional structures of macromolecules essential for drug discovery. With advancements in artificial intelligence (AI) and cryoEM, the Protein Data Bank (PDB) is solidifying its role as a key resource for 3D macromolecular structures. These developments underscore the growing need for enhanced quality metrics and robust validation standards for experimental structures.
This review examines recent advancements in cryoEM for drug discovery, analyzing structure quality metrics, resolution improvements, metal-ligand and water molecule identification, and refinement software. It compares cryoEM with other techniques like XRC and NMR, emphasizing the global expansion of cryoEM facilities and its increasing significance in drug discovery.
CryoEM is revolutionizing structural biology and drug discovery, particularly for large, complex structures in induced proximity and antibody-antigen interactions. It supports vaccine design, CAR T-cell optimization, gene editing, and gene therapy. Combined with AI, cryoEM enhances particle identification and 3D structure determination. With recent breakthroughs, cryoEM is emerging as a crucial tool in drug discovery, driving the development of new, effective therapies.
大分子X射线晶体学（XRC）、核磁共振（NMR）和冷冻电子显微镜（cryoEM）是确定药物发现中至关重要的大分子原子水平三维结构的主要技术。随着人工智能（AI）和冷冻电子显微镜技术的进步，蛋白质数据库（PDB）正巩固其作为三维大分子结构关键资源的作用。这些进展凸显了对提高实验结构质量指标和强大验证标准的需求日益增长。
本综述考察了冷冻电子显微镜在药物发现方面的最新进展，分析了结构质量指标、分辨率提升、金属配体和水分子识别以及精修软件。它将冷冻电子显微镜与X射线晶体学和核磁共振等其他技术进行了比较，强调了冷冻电子显微镜设施的全球扩展及其在药物发现中日益重要的地位。
冷冻电子显微镜正在彻底改变结构生物学和药物发现，特别是对于诱导接近和抗体-抗原相互作用中的大型复杂结构。它支持疫苗设计、嵌合抗原受体（CAR）T细胞优化、基因编辑和基因治疗。与人工智能相结合，冷冻电子显微镜增强了颗粒识别和三维结构确定。随着最近的突破，冷冻电子显微镜正在成为药物发现中的关键工具，推动新的有效疗法的开发。
