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Luciferase-Induced Photouncaging: Bioluminolysis.
Journal article

Luciferase-Induced Photouncaging: Bioluminolysis.

  • Chang D School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, 30 quai Ernest-Ansermet, Geneva, Switzerland.
  • Lindberg E School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, 30 quai Ernest-Ansermet, Geneva, Switzerland.
  • Feng S School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, 30 quai Ernest-Ansermet, Geneva, Switzerland.
  • Angerani S School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, 30 quai Ernest-Ansermet, Geneva, Switzerland.
  • Riezman H School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, 30 quai Ernest-Ansermet, Geneva, Switzerland.
  • Winssinger N School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, 30 quai Ernest-Ansermet, Geneva, Switzerland.
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  • 2019-09-04
Published in:
  • Angewandte Chemie (International ed. in English). - 2019
BRET
halotag
luciferase
optochemical reactions
uncaging reactions
Coumarins
Fluorescence Resonance Energy Transfer
HeLa Cells
Humans
Luciferases
Luminescent Measurements
Luminescent Proteins
Protein Binding
Pyrazoles
Pyrimidines
English Bioluminescence resonance energy transfer (BRET) has been widely used for studying dynamic processes in biological systems such as protein-protein interactions and other signaling events. Aside from acting as a reporter, BRET can also turn on functions in living systems. Herein, we report the application of BRET to performing a biorthogonal reaction in living cells; namely, releasing functional molecules through energy transfer to a coumarin molecule, a process termed bioluminolysis. An efficient BRET from Nanoluc-Halotag chimera protein (H-Luc) to a coumarin substrate yields the excited state of coumarin, which in turn triggers hydrolysis to uncage a target molecule. Compared to the conventional methods, this novel uncaging system requires no external light source and shows fast kinetics (t1/2 <2 min). We applied this BRET uncaging system to release a potent kinase inhibitor, ibrutinib, in living cells, highlighting its broad utility in controlling the supply of bioactive small molecules in vivo.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/242738
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