title: Thioflavin T as a fluorescence probe for label-free detection of T4 polynucleotide kinase/phosphatase and its inhibitors.
abstract: We have developed a new methodology for label-free fluorescence turn-on detection of T4 polynucleotide kinase/phosphatase activity (T4 PNKP) using a Thioflavin T probe. This method is very sensitive with a 0.01 unit/mL limit of detection, which is better than those with labeled fluorophores. Furthermore, T4 PNKP inhibition by the inhibitor heparin is shown, demonstrating the potential to screen suitable inhibitor drugs for T4 PNKP.
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China; State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, 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: Shunxin
authors.initials: S
authors.lastname: Jin
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.
authors.forename: Kun
authors.initials: K
authors.lastname: Xia
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Tang
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410081, China.
authors.forename: Kemin
authors.initials: K
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Medical Genetics & School of Life Sciences, Central South University, Changsha 410013, China. Electronic address: junwang@csu.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
mesh_terms: D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D017122:Bacteriophage T4 / Q000201:enzymology*; D052160:Benzothiazoles; D004791:Enzyme Inhibitors / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D052160:Benzothiazoles; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D013844:Thiazoles; C009462:thioflavin T; D011116:Polynucleotide 5'-Hydroxyl-Kinase
keywords: Label-free; Polynucleotide kinase/phosphatase; Thioflavin T