Kostyuk Alexander I, Rapota Diana D, Morozova Kseniia I, Fedotova Anna A, Jappy David, Semyanov Alexey V, Belousov Vsevolod V, Brazhe Nadezda A, Bilan Dmitry S
M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia; Pirogov Russian National Research Medical University, 117997, Moscow, Russia.
M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia.
Free Radic Biol Med. 2024 May 1;217:68-115. doi: 10.1016/j.freeradbiomed.2024.03.010. Epub 2024 Mar 18.
The objective of the current review is to summarize the current state of optical methods in redox biology. It consists of two parts, the first is dedicated to genetically encoded fluorescent indicators and the second to Raman spectroscopy. In the first part, we provide a detailed classification of the currently available redox biosensors based on their target analytes. We thoroughly discuss the main architecture types of these proteins, the underlying engineering strategies for their development, the biochemical properties of existing tools and their advantages and disadvantages from a practical point of view. Particular attention is paid to fluorescence lifetime imaging microscopy as a possible readout technique, since it is less prone to certain artifacts than traditional intensiometric measurements. In the second part, the characteristic Raman peaks of the most important redox intermediates are listed, and examples of how this knowledge can be implemented in biological studies are given. This part covers such fields as estimation of the redox states and concentrations of Fe-S clusters, cytochromes, other heme-containing proteins, oxidative derivatives of thiols, lipids, and nucleotides. Finally, we touch on the issue of multiparameter imaging, in which biosensors are combined with other visualization methods for simultaneous assessment of several cellular parameters.
本综述的目的是总结氧化还原生物学中光学方法的现状。它由两部分组成，第一部分致力于基因编码荧光指示剂，第二部分致力于拉曼光谱。在第一部分中，我们根据其目标分析物对目前可用的氧化还原生物传感器进行了详细分类。我们深入讨论了这些蛋白质的主要结构类型、其开发的基础工程策略、现有工具的生化特性以及从实际角度来看它们的优缺点。特别关注荧光寿命成像显微镜作为一种可能的读出技术，因为它比传统的强度测量更不容易出现某些伪影。在第二部分中，列出了最重要的氧化还原中间体的特征拉曼峰，并给出了如何将这些知识应用于生物学研究的示例。这部分涵盖了诸如铁硫簇、细胞色素、其他含血红素蛋白质、硫醇、脂质和核苷酸的氧化衍生物的氧化还原状态和浓度估计等领域。最后，我们涉及多参数成像问题，其中生物传感器与其他可视化方法相结合，用于同时评估几个细胞参数。
