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Robustness of NanoBiT luciferase complementation technology in the presence of widely used kinase inhibitors

Lookup NU author(s): Dr Tyrell Cartwright, Dr Stephanie Meyer, Professor Jonathan HigginsORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

Bioluminescence assays using luciferase enzymes are widely used in research to monitor gene expression and an array of other cell properties, and split luciferase enzymes can be used to measure protein interactions in biochemical assays and in living cells. When these methods are employed in chemical library screening efforts, it is vital that the activity of the luciferase enzyme itself is not strongly influenced by library components. Here, we developed a NanoBiT split luciferase assay to measure phosphorylation of Histone H3 peptides and used it to test the robustness of split luciferase to interference from two libraries of commonly used kinase inhibitors, including the Kinase Chemogenomic Set (KCGS). We found that NanoBiT luciferase is not significantly affected by the great majority of kinase inhibitors tested. However, the weak inhibition observed for a small minority of kinase inhibitors encourages the inclusion of suitable controls in NanoBiT (or NanoLuc) assays.

Publication metadata

Author(s): Cartwright TN, Meyer SK, Higgins JMG

Publication type: Article

Publication status: Published

Journal: SLAS Discovery

Year: 2022

Volume: 27

Issue: 8

Pages: 471-475

Print publication date: 03/12/2022

Online publication date: 23/09/2022

Acceptance date: 21/09/2022

Date deposited: 31/10/2022

ISSN (print): 2472-5552

ISSN (electronic): 2472-5560

Publisher: Elsevier

URL: https://doi.org/10.1016/j.slasd.2022.09.004

DOI: 10.1016/j.slasd.2022.09.004

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