期刊
BIOSENSORS & BIOELECTRONICS
卷 66, 期 -, 页码 77-83出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2014.11.014
关键词
Protein kinase; Carboxypeptidase Y; Phosphorylation; Electrochemical biosensor
类别
资金
- National Natural Science Foundation of China [21105056, 21375079]
- China Postdoctoral Science Foundation [2014M550369]
- Project of Development of Science and Technology of Shandong Province, China [2013GZX20109]
An effective assay method for monitoring protein kinase activity and screening inhibitors is greatly beneficial to kinase-related drug discovery, early diagnosis of diseases, and therapeutic effect evaluation. Herein, we develop a simple electrochemical method for detecting the activity of casein kinase II (CK2) based on phosphorylation against carboxypeptidase Y (CPY) digestion triggered signal amplification, where CK2 catalyzed phosphorylation event protects the substrate peptide from the digestion of CPY, maintains the repulsive force of the substrate peptide towards the redox probe, and results in a weak electrochemical signal. Whereas, without phosphorylation, the substrate peptide is digested by CPY and a strong electrochemical signal is obtained. The detection feasibility is demonstrated for the assay of CK2 activity with low detection limit of 0.047 unit/mL. Moreover, the biosensor was used for the analysis of kinase inhibition. Based on the electrochemical signal dependent inhibitor concentration, the IC50 value of ellagic acid was estimated to be 39.77 nM. The proposed method is also successfully applied to analyze CK2 activity in cell lysates, proving the applicability in complex biological samples. (C) 2014 Elsevier B.V. All rights reserved.
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