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Updated: Apr 26, 2026
Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
A mix-and-read fluorescence strategy for the switch-on probing of kinase activity based on an
Chunyang Lei1, Xiahong Xu, Jiang Zhou
1State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082 (P. R. China), Fax: (+86) 731-8882-1848.
Abstract:
Protein kinase plays a vital role in regulating signal-transduction pathways and its simple and quick detection is highly desirable because traditional kinase assays typically rely on a time-consuming kinase-phosphorylation process (ca. 1 h). Herein, we report a new and rapid fluorescence-based sensing platform for probing the activity of protein kinase that is based on the super-quenching capacity of graphene oxide (GO) nanosheets and specific recognition of the aptameric peptide (FITC-IP20). On the GO/peptide platform, the fluorescence quenching of FITC-IP20 that is adsorbed onto GO can be restored by selective binding of active protein kinase to the aptameric peptide, thereby resulting in the fast switch-on detection of kinase activity (ca. 15 min). The feasibility of this method has been demonstrated by the sensitive measurement of the activity of cAMP-dependent protein kinase (PKA), with a detection limit of 0.053 mU μL(-1). This assay technique was also successfully applied to the detection of kinase activation in cell lysate.
