To tilt or not to tilt? Strategies for in situ cryo-EM data collection

IF 6.1 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Current opinion in structural biology Pub Date : 2025-06-28 DOI:10.1016/j.sbi.2025.103100

Joshua L. Dickerson , Bronwyn A. Lucas

引用次数: 0

Abstract

Recent breakthroughs in single-particle cryogenic electron microscopy (cryo-EM) and protein structure prediction have transformed our ability to resolve molecular structures. Since we have now experimentally determined, or can confidently predict, the structures of a significant portion of the proteome, and since workflows for imaging in cells are established, the stage is set for applying cryo-EM to understand the molecular basis of complex cellular functions. This review explores a spectrum of data collection strategies—from 2D approaches to tomography—used for in situ cryo-EM. We discuss their relative merits in addressing key biological questions and the need to tailor them towards experimental goals. Improvements in theoretical and practical understanding of the challenges for in situ cryo-EM are necessary for optimizing data collection strategies and pushing the boundaries of structural cell biology.

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倾斜还是不倾斜？原位低温电镜数据收集策略

最近在单粒子低温电子显微镜（cryo-EM）和蛋白质结构预测方面的突破改变了我们解决分子结构的能力。由于我们现在已经通过实验确定或可以自信地预测蛋白质组的重要部分的结构，并且由于在细胞中成像的工作流程已经建立，因此应用冷冻电镜来理解复杂细胞功能的分子基础的阶段已经确定。这篇综述探讨了一系列数据收集策略-从二维方法到层析成像-用于原位冷冻电镜。我们讨论了它们在解决关键生物学问题方面的相对优点，以及对实验目标进行调整的必要性。改进对原位冷冻电镜挑战的理论和实践理解对于优化数据收集策略和推动结构细胞生物学的界限是必要的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current opinion in structural biology 生物-生化与分子生物学

CiteScore

12.20

自引率

2.90%

发文量

179

审稿时长

6-12 weeks

期刊介绍： Current Opinion in Structural Biology (COSB) aims to stimulate scientifically grounded, interdisciplinary, multi-scale debate and exchange of ideas. It contains polished, concise and timely reviews and opinions, with particular emphasis on those articles published in the past two years. In addition to describing recent trends, the authors are encouraged to give their subjective opinion of the topics discussed. In COSB, we help the reader by providing in a systematic manner: 1. The views of experts on current advances in their field in a clear and readable form. 2. Evaluations of the most interesting papers, annotated by experts, from the great wealth of original publications. [...] The subject of Structural Biology is divided into twelve themed sections, each of which is reviewed once a year. Each issue contains two sections, and the amount of space devoted to each section is related to its importance. -Folding and Binding- Nucleic acids and their protein complexes- Macromolecular Machines- Theory and Simulation- Sequences and Topology- New constructs and expression of proteins- Membranes- Engineering and Design- Carbohydrate-protein interactions and glycosylation- Biophysical and molecular biological methods- Multi-protein assemblies in signalling- Catalysis and Regulation