Fluorophores have transformed the way we study biological systems, enabling non-invasive studies in cells and intact organisms, which increase our understanding of complex processes at the molecular level. Fluorescent amino acids have become an essential chemical tool because they can be used to construct fluorescent macromolecules, such as peptides and proteins, without disrupting their native biomolecular properties. Fluorescent and fluorogenic amino acids with unique photophysical properties have been designed for tracking protein–protein interactions in situ or imaging nanoscopic events in real time with high spatial resolution. In this Review, we discuss advances in the design and synthesis of fluorescent amino acids and how they have contributed to the field of chemical biology in the past 10 years. Important areas of research that we review include novel methodologies to synthesize building blocks with tunable spectral properties, their integration into peptide and protein scaffolds using site-specific genetic encoding and bioorthogonal approaches, and their application to design novel artificial proteins, as well as to investigate biological processes in cells by means of optical imaging.

荧光团改变了我们研究生物系统的方式，使细胞和完整生物体的非侵入性研究成为可能，这增加了我们在分子水平上对复杂过程的理解。荧光氨基酸已成为一种重要的化学工具，因为它们可用于构建荧光大分子，如肽和蛋白质，而不会破坏其天然生物分子特性。具有独特光物理特性的荧光和荧光氨基酸已被设计用于原位跟踪蛋白质-蛋白质相互作用或以高空间分辨率实时成像纳米级事件。在这篇综述中，我们讨论了荧光氨基酸设计和合成的进展，以及它们在过去 10 年中如何为化学生物学领域做出贡献。