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Quantification of ERK Kinase Activity in Biological Samples Using Differential Sensing
Diana Zamora-Olivares1,2, Tamer S Kaoud3,4, Lingyu Zeng5
1Department of Chemistry , The University of Texas at Austin , Austin , Texas 78712 , United States.
ACS Chemical Biology
|November 28, 2019
Summary
A new biosensor array rapidly quantifies active protein kinase activity in cell samples. This method offers a high-throughput, reproducible alternative to traditional techniques for studying cell signaling networks.
Area of Science:
- Biochemistry
- Molecular Biology
- Systems Biology
Background:
- Understanding complex biological systems necessitates evaluating gene, protein, and reaction networks.
- Protein phosphorylation is crucial for cell signaling, but current quantification methods like Western blots and proteomics have limitations in reproducibility and direct activity correlation.
- There is a need for technologies enabling rapid and quantitative assessment of biological networks, particularly protein kinase activity.
Purpose of the Study:
- To present a novel, complementary approach for the rapid quantification of protein kinase activity in biological samples.
- To demonstrate the utility of this approach using the activity of extracellular signal-regulated kinase (ERK) as a model.
- To highlight the potential for high-throughput analysis and quantification of multiple kinase activities in a single sample.
Main Methods:
- Development of an array sensing approach utilizing a library of differential peptide-based biosensors.
- Application of chemometric tools for data analysis.
- Testing the method on cell lysates and tumor extracts to quantify active ERK.
Main Results:
- Successfully quantified nanograms of active ERK in micrograms of unfractionated cell lysates and tumor extracts.
- Demonstrated the ability to rapidly measure kinase activity without the need for exogenous inhibitors.
- Showcased the potential for quantifying multiple protein kinase activities simultaneously.
Conclusions:
- The presented differential sensing approach offers a rapid, reproducible, and quantitative method for assessing protein kinase activity.
- This technology can significantly advance the development of biological models, identification of treatment targets, and assessment of treatment plans.
- The method overcomes limitations of traditional techniques by directly quantifying active kinase levels, facilitating high-throughput biological network analysis.
