Ivanova Irina A, Valueva Anastasia A, Ershova Maria O, Pleshakova Tatiana O
Institute of Biomedical Chemistry, Pogodinskaya Str., 10, 119121 Moscow, Russia.
Biomolecules. 2025 Apr 12;15(4):574. doi: 10.3390/biom15040574.
The conventional approach to investigating enzyme systems involves the simultaneous investigation of a large number of molecules and observing ensemble-averaged properties. However, modern science allows us to study the properties of single molecules and to obtain data on biochemical systems at a fundamentally new level, significantly expanding our understanding of the mechanisms of biochemical processes. Imaging of single biomolecules with high spatial and temporal resolution is among such modern research tools. To effectively image the individual steps or intermediates of biochemical reactions in single-molecule experiments, we need to develop a methodology for data acquisition and analysis. Its development will make it possible to solve the problem of separating the static and dynamic disorder present in the parameters identified by traditional proteomic methods. Such a methodology may be based on AFM imaging, the high-resolution microscopic visualization of enzymes. This review focuses on this direction of research, including the relevant methodological and practical solutions related to the potential of developing a single-molecule approach to the study of enzyme systems using AFM-based techniques. We focus on the results of enzyme reaction studies, as there are still few such studies, as opposed to the AFM studies of the mechanical properties of individual enzyme molecules.
研究酶系统的传统方法涉及同时研究大量分子并观察总体平均性质。然而，现代科学使我们能够研究单个分子的性质，并在一个全新的层面上获取有关生物化学系统的数据，极大地扩展了我们对生物化学过程机制的理解。具有高空间和时间分辨率的单生物分子成像就是这类现代研究工具之一。为了在单分子实验中有效地对生化反应的各个步骤或中间体进行成像，我们需要开发一种数据采集和分析方法。它的发展将有可能解决分离传统蛋白质组学方法所确定参数中存在的静态和动态无序问题。这样一种方法可能基于原子力显微镜（AFM）成像，即酶的高分辨率微观可视化。本综述聚焦于这一研究方向，包括与利用基于AFM的技术开发研究酶系统的单分子方法的潜力相关的方法和实际解决方案。我们关注酶反应研究的结果，因为与对单个酶分子力学性质的AFM研究相比，这类研究仍然很少。
