Multiple and cooperative binding of fluorescence light-up probe Thioflavin T with human telomere DNA G-quadruplex

Valérie Gabelica; Ryuichi Maeda; Takeshi Fujimoto; Hidenobu Yaku; Takashi Murashima; Naoki Sugimoto; Daisuke Miyoshi

doi:10.1021/bi4006072

Multiple and cooperative binding of fluorescence light-up probe Thioflavin T with human telomere DNA G-quadruplex

Valérie Gabelica ^{1, 2, *} Ryuichi Maeda ³ Takeshi Fujimoto ³ Hidenobu Yaku ^{3, 4, 5} Takashi Murashima ^{3, 4} Naoki Sugimoto ^{3, 4} Daisuke Miyoshi ^{3, 4, *}

* Auteur correspondant

1 ARNA - Régulations Naturelles et Artificielles

2 Department of Chemistry [Liège, Belgium]

3 FIRST - Faculty of Frontiers of Innovative Research in Science and Technology [Kobe, Japan]

4 FIBER - Frontier Institute for Biomol ecular Engineering Research [Kobe, Japan]

5 Advanced Technology Research Laboratories [Kyoto, Japan]

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.

Keywords : G-quadruplex DNA G-quadruplex ligands Mass spectrometry Native mass spectrometry

Type de document :

Article dans une revue

Biochemistry, American Chemical Society, 2013, 52, pp.5620 - 5628. 〈10.1021/bi4006072 〉

Domaine :

Sciences du Vivant [q-bio] / Biochimie, Biologie Moléculaire

Liste complète des métadonnées

http://www.hal.inserm.fr/inserm-01388017
Contributeur : Valérie Gabelica <>
Soumis le : mercredi 26 octobre 2016 - 13:02:04
Dernière modification le : vendredi 28 octobre 2016 - 16:46:29

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Multiple and cooperative binding of fluorescence light-up probe Thioflavin T with human telomere DNA G-quadruplex

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