title: G-quadruplex-based fluorometric biosensor for label-free and homogenous detection of protein acetylation-related enzymes activities.
abstract: Reversible protein acetylation, one of the key types of post-translational modifications, is composed of histone acetylation and deacetylation, which is typically catalyzed by histone acetyltransferases (HATs) and histone deacetylases (HDACs) respectively. Herein, a label-free fluorescent method has been established for the homogeneous bioassay of HAT/HDAC activity and respective inhibitors. The proposed approach is primarily based on the electrostatic interaction between G-quadruplexes (G4s) and acetylation-related peptides, which results in marked change of fluorescent intensity of G4/Thioflavin T (ThT) complexes. This HAT (p300) activity assay is exceedingly sensitive and selective, with a linear range from 0.1 to 120nM and a detection limit of 0.05nM. Moreover, this biosensor is feasible to detect the HDAC (Sirt1) activity with a linear range from 1 to 450nM and a detection limit of 1nM. The potency of this assay is further demonstrated by detecting HAT/HDAC activity in cell lysates and evaluating HAT and HDAC-targeted inhibitors, C464 and EX 527, respectively. The proposed assay is convenient, label-free and cost-efficient, which is promising for HAT/HDAC-targeted epigenetic research and pharmaceutical development.
authors.affiliation: State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, People's Republic of China.
authors.forename: Huixia
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, People's Republic of China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Li
authors.affiliation: State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, People's Republic of China.
authors.forename: Kunli
authors.initials: K
authors.lastname: Zhao
authors.affiliation: State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, People's Republic of China.
authors.forename: Siyi
authors.initials: S
authors.lastname: Chen
authors.affiliation: State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, People's Republic of China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Medicinal Chemistry, School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, People's Republic of China; Key Laboratory of Structure-Based Drug Design & Discovery, Ministry of Education, Shenyang Pharmaceutical University, Shenyang 110016, People's Republic of China. Electronic address: randybinlin@gmail.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Lin
authors.affiliation: State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, People's Republic of China. Electronic address: niezhou.hnu@gmail.com.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Nie
authors.affiliation: State Key laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, People's Republic of China.
authors.forename: Shouzhuo
authors.initials: S
authors.lastname: Yao
mesh_terms: D000107:Acetylation; D052160:Benzothiazoles; D015374:Biosensing Techniques / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057075:Enzyme Assays / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry*; D054856:G-Quadruplexes; D006367:HeLa Cells; D051548:Histone Acetyltransferases / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D057230:Limit of Detection; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013844:Thiazoles / Q000737:chemistry*
chemical_list: D052160:Benzothiazoles; D005456:Fluorescent Dyes; D056572:Histone Deacetylase Inhibitors; D010455:Peptides; D013844:Thiazoles; C009462:thioflavin T; D051548:Histone Acetyltransferases; D006655:Histone Deacetylases
keywords: G-quadruplexes; Histone acetyltransferases; Histone deacetylases; Label-free fluorescent method; Thioflavin T