荧光素酶诱导的小分子效应物光还原解笼。

Department of Organic Chemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, 30 quai Ernest Ansermet, 1211, Geneva, Switzerland.

Nat Commun. 2018 Aug 30;9(1):3539. doi: 10.1038/s41467-018-05916-9.

Bioluminescence resonance energy transfer (BRET) is extensively used to study dynamic systems and has been utilized in sensors for studying protein proximity, metabolites, and drug concentrations. Herein, we demonstrate that BRET can activate a ruthenium-based photocatalyst which performs bioorthogonal reactions. BRET from luciferase to the ruthenium photocatalyst is used to uncage effector molecules with up to 64 turnovers of the catalyst, achieving concentrations >0.6 μM effector with 10 nM luciferase construct. Using a BRET sensor, we further demonstrate that the catalysis can be modulated in response to an analyte, analogous to allosterically controlled enzymes. The BRET-induced reaction is used to uncage small-molecule drugs (ibrutinib and duocarmycin) at biologically effective concentrations in cellulo.

生物发光共振能量转移（BRET）被广泛用于研究动态系统，并已被用于研究蛋白质接近度、代谢物和药物浓度的传感器中。在这里，我们证明 BRET 可以激活基于钌的光催化剂，该催化剂可进行生物正交反应。荧光素酶到钌光催化剂的 BRET 用于释放具有高达 64 个催化剂周转率的效应分子，实现了具有 10 nM 荧光素酶构建体的 >0.6 μM 效应物浓度。使用 BRET 传感器，我们进一步证明可以响应分析物来调节催化作用，类似于变构控制的酶。BRET 诱导的反应用于在细胞内以生物有效浓度释放小分子药物（依鲁替尼和 duocarmycin）。