Related Experiment Video
Updated: Mar 7, 2026
Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
Published on: June 30, 2019
One reporter for in-cell activity profiling of majority of protein kinase oncogenes
Iva Gudernova1, Silvie Foldynova-Trantirkova2, Barbora El Ghannamova2
1Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
Abstract:
In-cell profiling enables the evaluation of receptor tyrosine activity in a complex environment of regulatory networks that affect signal initiation, propagation and feedback. We used FGF-receptor signaling to identify EGR1 as a locus that strongly responds to the activation of a majority of the recognized protein kinase oncogenes, including 30 receptor tyrosine kinases and 154 of their disease-associated mutants. The EGR1 promoter was engineered to enhance trans-activation capacity and optimized for simple screening assays with luciferase or fluorescent reporters. The efficacy of the developed, fully synthetic reporters was demonstrated by the identification of novel targets for two clinically used tyrosine kinase inhibitors, nilotinib and osimertinib. A universal reporter system for in-cell protein kinase profiling will facilitate repurposing of existing anti-cancer drugs and identification of novel inhibitors in high-throughput screening studies.
Insights
Researchers developed a universal reporter system for in-cell protein kinase profiling. This system identifies novel drug targets and facilitates repurposing of anti-cancer drugs for high-throughput screening.
Area of Science:
- Molecular Biology
- Cell Signaling
- Oncology
Background:
- Receptor tyrosine kinase (RTK) activity is crucial in cellular signaling and often dysregulated in cancer.
- Evaluating RTK activity within complex cellular environments is challenging.
- Understanding RTK signaling networks is vital for developing targeted cancer therapies.
Purpose of the Study:
- To develop a universal reporter system for in-cell protein kinase profiling.
- To identify novel targets for kinase inhibitors.
- To facilitate drug repurposing and high-throughput screening for anti-cancer agents.
Main Methods:
- Engineered the *EGR1* promoter for enhanced trans-activation capacity.
- Developed synthetic reporters using luciferase or fluorescent readouts.
- Utilized FGF-receptor signaling to identify *EGR1* as a responsive locus.
- Tested reporter efficacy by identifying novel targets for nilotinib and osimertinib.
Main Results:
- Identified *EGR1* as a locus responsive to the activation of numerous protein kinase oncogenes, including 30 RTKs and 154 disease-associated mutants.
- Demonstrated the efficacy of the synthetic reporter system in identifying novel targets for tyrosine kinase inhibitors.
- Successfully identified new targets for nilotinib and osimertinib.
Conclusions:
- A universal reporter system for in-cell protein kinase profiling has been successfully developed.
- This system enables efficient evaluation of kinase activity in complex cellular networks.
- The reporter system will accelerate drug repurposing and the discovery of novel anti-cancer inhibitors through high-throughput screening.
Related Concept Videos
01:22PI3K/mTOR/AKT Signaling Pathway
02:54Protein Kinases and Phosphatases
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
02:54Protein Kinases and Phosphatases
03:03mTOR Signaling and Cancer Progression
The mTOR pathway or the...
01:19Interactions Between Signaling Pathways
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
02:34Inhibition of Cdk Activity