Novel BRET Combination for Detection of Rapamycin-induced Protein Dimerization Using Luciferase from Fungus

Overview

Journal Heliyon

Specialty Social Sciences

Date 2024 Feb 22

PMID 38384550

Authors

Aaiyas Mujawar

Shalini Dimri

Ksenia A Palkina

Nadezhda M Markina

Karen S Sarkisyan

Anastasia V Balakireva

Ilia V Yampolsky

Abhijit De

Affiliations

Soon will be listed here.

Abstract

Bioluminescence resonance energy transfer (BRET) is one of the most promising approaches used for noninvasive imaging of protein-protein interactions . Recently, our team has discovered a genetically encodable bioluminescent system from the fungus and identified a novel luciferase that represents an imaging tool orthogonal to other luciferin-luciferase systems. We demonstrated the possibility of using the fungal luciferase as a new BRET donor by creating fused pairs with acceptor red fluorescent proteins, of which tdTomato provided the highest BRET efficiency. Using this new BRET system, we also designed a mTOR pathway specific rapamycin biosensor by integrating the FRB and FKBP12 protein dimerization system. We demonstrated the specificity and efficacy of the new fungal luciferase-based BRET combination for application in mammalian cell culture that will provide the unique opportunity to perform multiplexed BRET assessment in the future.

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