title: A mix-and-read fluorescence strategy for the switch-on probing of kinase activity based on an aptameric-peptide/graphene-oxide platform.
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.
authors.affiliation: State 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.
authors.forename: Chunyang
authors.initials: C
authors.lastname: Lei
authors.forename: Xiahong
authors.initials: X
authors.lastname: Xu
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhou
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.forename: Zhou
authors.initials: Z
authors.lastname: Nie
authors.forename: Meng
authors.initials: M
authors.lastname: Qing
authors.forename: Pei
authors.initials: P
authors.lastname: Li
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.forename: Shouzhuo
authors.initials: S
authors.lastname: Yao
mesh_terms: D052158:Aptamers, Peptide / Q000737:chemistry / Q000378:metabolism*; D015374:Biosensing Techniques; D004789:Enzyme Activation; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006108:Graphite / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D061986:MCF-7 Cells; D010087:Oxides / Q000737:chemistry / Q000378:metabolism*; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014407:Tumor Cells, Cultured
chemical_list: D052158:Aptamers, Peptide; D005456:Fluorescent Dyes; D010087:Oxides; D006108:Graphite; D011494:Protein Kinases
keywords: biosensors; fluorescence; graphene; kinase; peptides