title: Multiple and cooperative binding of fluorescence light-up probe thioflavin T with human telomere DNA G-quadruplex.
abstract: Thioflavin T (ThT), a typical probe for protein fibrils, also binds human telomeric G-quadruplexes with a fluorescent light-up signal change and high specificity against DNA duplexes. Cell penetration and low cytotoxicity of fibril probes having been widely established, modifying ThT and other fibril probes is an attractive means of generating new G-quadruplex ligands. Thus, elucidating the binding mechanism is important for the design of new drugs and fluorescent probes based on ThT. Here, we investigated the binding mechanism of ThT with several variants of the human telomeric sequence in the presence of monovalent cations. Fluorescence titrations and electrospray ionization mass spectrometry (ESI-MS) analyses demonstrated that each G-quadruplex unit cooperatively binds to several ThT molecules. ThT brightly fluoresces when a single ligand is bound to the G-quadruplex and is quenched as ligand binding stoichiometry increases. Both the light-up signal and the dissociation constants are exquisitely sensitive to the base sequence and to the G-quadruplex structure. These results are crucial for the sensible design and interpretation of G-quadruplex detection assays using fluorescent ligands in general and ThT in particular.
authors.affiliation: Physical Chemistry and Mass Spectrometry Laboratory, Department of Chemistry, University of Liège, B-4000 Liège, Belgium. v.gabelica@iecb.u-bordeaux.fr
authors.forename: Valérie
authors.initials: V
authors.lastname: Gabelica
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Maeda
authors.forename: Takeshi
authors.initials: T
authors.lastname: Fujimoto
authors.forename: Hidenobu
authors.initials: H
authors.lastname: Yaku
authors.forename: Takashi
authors.initials: T
authors.lastname: Murashima
authors.forename: Naoki
authors.initials: N
authors.lastname: Sugimoto
authors.forename: Daisuke
authors.initials: D
authors.lastname: Miyoshi
mesh_terms: D000465:Algorithms; D052160:Benzothiazoles; D001665:Binding Sites; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D054856:G-Quadruplexes; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D015394:Molecular Structure; D021241:Spectrometry, Mass, Electrospray Ionization; D016615:Telomere / Q000235:genetics*; D013844:Thiazoles / Q000737:chemistry* / Q000378:metabolism
chemical_list: D052160:Benzothiazoles; D005456:Fluorescent Dyes; D013844:Thiazoles; C009462:thioflavin T; D004247:DNA