title: Label-free colorimetric assay for T4 polynucleotide kinase/phosphatase activity and its inhibitors based on G-quadruplex/hemin DNAzyme.
abstract: Here we report a new approach for label-free colorimetric assay of T4 polynucleotide kinase/phosphatase (PNKP) activity based on G-quadruplex/hemin DNAzyme. In the presence of T4 PNKP, the DNA primer with a 3'-phosphate can be dephosphorylated into a 3'-hydroxyl and initiate a primer elongation reaction to open the hairpin probe, and leading to releasing the G-quartets. Under optimal conditions, the proposed method exhibited a considerable performance with a detection limit of 0.01 U/mL. Furthermore, the present assay can be used to study the potential T4 PNKP inhibitor screening, making it promise to be applied in the fields of drug discovery.
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors.forename: Haisheng
authors.initials: H
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China. Electronic address: macb2012@csu.edu.cn.
authors.forename: Changbei
authors.initials: C
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors.forename: Hanchun
authors.initials: H
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, China.
authors.forename: Kemin
authors.initials: K
authors.lastname: Wang
mesh_terms: D017122:Bacteriophage T4 / Q000201:enzymology*; D003124:Colorimetry / Q000379:methods; D021881:DNA, Catalytic / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000737:chemistry*; D054856:G-Quadruplexes; D006427:Hemin / Q000737:chemistry*; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000032:analysis*; D014764:Viral Proteins / Q000032:analysis*
chemical_list: D021881:DNA, Catalytic; D004791:Enzyme Inhibitors; D014764:Viral Proteins; D006427:Hemin; D011116:Polynucleotide 5'-Hydroxyl-Kinase
keywords: Colorimetric assay; G-quadruplex/hemin DNAzyme; T4 polynucleotide kinase/phosphatase