Luciferase-induced photoreductive uncaging of small-molecule effectors

Abstract

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.

Luciferase‐Induced Photouncaging: Bioluminolysis

“…BRET has been utilized to control neuronal activity, gene expression, and a photo‐switchable fluorescent protein . A BRET‐induced chemical reaction can also be used to change the function of bioactive molecules, and we reported a BRET‐induced ruthenium‐photocatalyzed reaction . While these precedents highlight novel applications, the extension of the scope of the chemistry that can be performed with bioluminescence is empowering.…”

Transition Metal‐Promoted Reactions in Aqueous Media and Biological Settings

“…The precise designo faluciferase-based indicator of drugs (LUCID), involving aS NAP protein, the luciferase and a receptor protein that binds the caged drug, is keyf or the success of the strategy,s ince the luciferase, ruthenium complex and the caged molecule need to be in close proximity. [133]…”

Visible Light Photoredox Catalysis Using Ruthenium Complexes in Chemical Biology

“…In parallel work, we have shown that ab ioluminescence resonancee nergy transfer (BRET) can be used to photoexcite the ruthenium and enable catalysis (Figure 10 D). [70] BRET from luciferase to the ruthenium photocatalyst was used to uncage different effector molecules (drugs or af luorophore) with up to 64 turnoverso ft he catalyst, achieving concentrations > 0.6 mm effector with 10 nm luciferase construct. Using aB RET sensor, we furtherd emonstrated that this catalysis could be modulated in response to an analyte, analogous to an allosterically controlled enzyme.…”