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Assaying Protein Kinase Activity with Radiolabeled ATP
Published on: May 26, 2017
A universal and multiplex kinase assay using γ-[(18)O(4)]-ATP
Chuan Fu1, Xueyun Zheng, Yao Jiang
1Department of Chemistry, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, PR China.
Chemical Communications (Cambridge, England)
|February 27, 2013
Summary
A novel non-radioactive method using γ-[(18)O4]-ATP enables multiplex kinase assays. This approach is broadly applicable, allowing for the assessment of inhibitor effectiveness and specificity across various kinases without specialized reagents.
Area of Science:
- Biochemistry
- Enzymology
- Chemical Biology
Background:
- Kinase activity assays are crucial for drug discovery and biological research.
- Current methods often rely on radioactive substrates or purified enzymes, limiting their scope and application.
- Developing non-radioactive, versatile assay platforms is a significant need in the field.
Purpose of the Study:
- To develop a novel, non-radioactive, and multiplex assay for kinase activity.
- To demonstrate the broad applicability of this method across different kinases.
- To enable the assessment of kinase inhibitor effectiveness and specificity.
Main Methods:
- Synthesis of γ-[(18)O4]-adenosine triphosphate (ATP).
- Development of a non-radioactive assay utilizing the synthesized ATP analog.
- Application of the assay to evaluate kinase activity and inhibitor profiles.
Main Results:
- Successful synthesis of γ-[(18)O4]-ATP.
- Establishment of a robust non-radioactive assay applicable to various kinases.
- Demonstration of the method's utility in determining inhibitor effectiveness and specificity without purified enzymes or fluorescent substrates.
Conclusions:
- The developed γ-[(18)O4]-ATP based method offers a versatile and non-radioactive approach for kinase research.
- This novel assay platform simplifies kinase inhibitor screening and characterization.
- The method's broad applicability and lack of requirement for specialized reagents make it a valuable tool for biochemical and pharmacological studies.
