期刊
ANALYTICA CHIMICA ACTA
卷 934, 期 -, 页码 36-43出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2016.06.024
关键词
Photoelectrochemical biosensor; Protein kinase activity; Phosphorylated graphite-like carbon nitride; Inhibitor; Real samples
资金
- National Natural Science Foundation of China [21375079]
- Natural Science Foundation of Shandong province, China [ZR2014BQ029]
- China Postdoctoral Science Foundation [2014M550369, 2015M582120]
- Project of Development of Science and Technology of Shandong Province, China [2013GZX20109]
Protein kinases are general and significant regulators in the cell signaling pathway, and it is still greatly desired to achieve simple and quick kinase detection. Herein, we develop a simple and sensitive photo-electrochemical strategy for the detection of protein kinase activity based on the bond between phosphorylated peptide and phosphorylated graphite-like carbon nitride (P-g-C3N4) conjugates triggered by Zr4+ ion coordination. Under optimal conditions, the increased photocurrent is proportional to the protein kinase A (PKA) concentration ranging from 0.05 to 50 U/mL with a detection limit of 0.077 U/mL. Moreover, this photoelectrochemical assay can be also applied to quantitative analysis of kinase inhibition. The results indicated that the IC50 value (inhibitor concentration producing 50% inhibitor) for ellagic acid was 9.1 mu M. Moreover, the developed method is further applied to detect PKA activity in real samples, which contains serum from healthy person and gastric cancer patients and breast tissue from healthy person and breast cancer patients. Therefore, the established protocol provides a new and simple tool for assay of kinase activity and its inhibitors with low cost and high sensitivity. (C) 2016 Elsevier B.V. All rights reserved.
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