Price Sky, Que Emily L
Department of Chemistry, The University of Texas at Austin, Austin, TX 78712, USA.
Department of Chemistry, The University of Texas at Austin, Austin, TX 78712, USA.
Curr Opin Chem Biol. 2024 Aug;81:102475. doi: 10.1016/j.cbpa.2024.102475. Epub 2024 Jun 8.
Metalloenzymes are essential to cellular function, and their overexpression or enhanced activation are potential therapeutic targets. However, the study of metalloenzymes in vitro presents various challenges, leading many to develop tools to study them in their native cellular environment. Small-molecule fluorescence probes are commonly used to monitor metalloenzyme function, activity, and distribution in situ. These include probes that are activity-based (fluorescence is mediated by enzyme activity) or binding-based (fluorescence is mediated by interactions with the enzyme upon binding its metal cofactor). We discuss recent innovations that overcome key design challenges, such as the rapid diffusion of activity-based probes, the difficulty of probing redox-active enzymes, the selectivity of binding-based probes, and the poor penetration depth of fluorescence, and describe novel applications of these tools.
金属酶对细胞功能至关重要，其过表达或激活增强是潜在的治疗靶点。然而，体外研究金属酶面临各种挑战，促使许多人开发在天然细胞环境中研究它们的工具。小分子荧光探针通常用于原位监测金属酶的功能、活性和分布。这些探针包括基于活性的（荧光由酶活性介导）或基于结合的（荧光由与酶结合其金属辅因子时的相互作用介导）。我们讨论了克服关键设计挑战的最新创新，例如基于活性的探针快速扩散、探测氧化还原活性酶的困难、基于结合的探针的选择性以及荧光穿透深度差等，并描述了这些工具的新应用。
