Fleming Cassandra L, Benitez-Martin Carlos, Bernson Elin, Xu Yongjin, Kristenson Linnea, Inghardt Tord, Lundbäck Thomas, Thorén Fredrik B, Grøtli Morten, Andréasson Joakim
Department of Chemistry and Chemical Engineering, Physical Chemistry, Chalmers University of Technology SE-41296 Göteborg Sweden
Department of Chemistry and Molecular Biology, University of Gothenburg Box 462 SE-40530 Göteborg Sweden
Chem Sci. 2024 Apr 12;15(18):6897-6905. doi: 10.1039/d4sc00390j. eCollection 2024 May 8.
Light-responsive molecular tools targeting kinases affords one the opportunity to study the underlying cellular function of selected kinases. In efforts to externally control lymphocyte-specific protein tyrosine kinase (LCK) activity, the development of release-and-report LCK inhibitors is described, in which (i) the release of the active kinase inhibitor can be controlled externally with light; and (ii) fluorescence is employed to report both the release and binding of the active kinase inhibitor. This introduces an unprecedented all-photonic method for users to both control and monitor real-time inhibitory activity. A functional cellular assay demonstrated light-mediated LCK inhibition in natural killer cells. The use of coumarin-derived caging groups resulted in rapid cellular uptake and non-specific intracellular localisation, while a BODIPY-derived caging group predominately localised in the cellular membrane. This concept of release-and-report inhibitors has the potential to be extended to other biorelevant targets where both spatiotemporal control in a cellular setting and a reporting mechanism would be beneficial.
靶向激酶的光响应分子工具为人们提供了研究特定激酶潜在细胞功能的机会。为了从外部控制淋巴细胞特异性蛋白酪氨酸激酶（LCK）的活性，本文描述了释放并报告型LCK抑制剂的研发，其中：（i）活性激酶抑制剂的释放可通过光从外部进行控制；（ii）利用荧光报告活性激酶抑制剂的释放和结合情况。这为用户引入了一种前所未有的全光学方法，用于控制和监测实时抑制活性。一项功能性细胞试验证明了在自然杀伤细胞中光介导的LCK抑制作用。使用香豆素衍生的笼蔽基团可实现快速的细胞摄取和非特异性的细胞内定位，而硼二吡咯衍生的笼蔽基团主要定位于细胞膜。这种释放并报告型抑制剂的概念有可能扩展到其他生物相关靶点，在细胞环境中进行时空控制和报告机制都将是有益的。
