title: A novel photoelectrochemical biosensor for protein kinase activity assay based on phosphorylated graphite-like carbon nitride.
abstract: 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 photoelectrochemical 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 Zr(4+) 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 μM. 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.
authors: College of Chemistry and Material Science, Shandong Agricultural University, Taian, 271018, PR China.
authors: Xue
authors: X
authors: Li
authors: College of Chemistry and Material Science, Shandong Agricultural University, Taian, 271018, PR China.
authors: Yunlei
authors: Y
authors: Zhou
authors: College of Chemistry and Material Science, Shandong Agricultural University, Taian, 271018, PR China.
authors: Yan
authors: Y
authors: Xu
authors: College of Chemistry and Material Science, Shandong Agricultural University, Taian, 271018, PR China.
authors: Huijie
authors: H
authors: Xu
authors: College of Chemistry and Material Science, Shandong Agricultural University, Taian, 271018, PR China.
authors: Minghui
authors: M
authors: Wang
authors: College of Chemistry and Material Science, Shandong Agricultural University, Taian, 271018, PR China. Electronic address: yinhs@sdau.edu.cn.
authors: Huanshun
authors: H
authors: Yin
authors: College of Chemistry and Material Science, Shandong Agricultural University, Taian, 271018, PR China. Electronic address: ashy@sdau.edu.cn.
authors: Shiyun
authors: S
authors: Ai
mesh_terms: D015374:Biosensing Techniques; D055664:Electrochemical Techniques; D004610:Ellagic Acid / Q000494:pharmacology; D006108:Graphite / Q000737:chemistry*; D009570:Nitriles / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D055668:Photochemical Processes; D011494:Protein Kinases / Q000032:analysis* / Q000378:metabolism; D015040:Zirconium / Q000737:chemistry / Q000378:metabolism
chemical_list: D009570:Nitriles; D004610:Ellagic Acid; C011206:cyanogen; D006108:Graphite; D015040:Zirconium; D011494:Protein Kinases
keywords: Inhibitor; Phosphorylated graphite-like carbon nitride; Photoelectrochemical biosensor; Protein kinase activity; Real samples