Article (Scientific journals)
Multiple and cooperative binding of fluorescence light-up probe thioflavin t with human telomere DNA G-quadruplex
Gabelica, Valerie; Maeda, R.; Fujimoto, T. et al.
2013In Biochemistry, 52 (33), p. 5620-5628
Peer Reviewed verified by ORBi
 

Files


Full Text
bi-2013-006072-author postprint.pdf
Author postprint (559 kB)
Download

All documents in ORBi are protected by a user license.

Send to



Details



Keywords :
Cooperative binding; Dissociation constant; Electrospray ionization mass spectrometry; Fluorescence titrations; Fluorescent probes; G-quadruplex structure; Human telomere DNA; Telomeric sequences; Biochemistry; Dissociation; Fluorescence; Ligands; Mass spectrometry; Probes; DNA sequences; DNA; DNA sequence; Algorithms; Binding Sites; Circular Dichroism; Fluorescent Dyes; G-Quadruplexes; Humans; Kinetics; Models, Molecular; Molecular Structure; Spectrometry, Mass, Electrospray Ionization; Telomere; Thiazoles
Abstract :
[en] 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. © 2013 American Chemical Society.
Disciplines :
Chemistry
Author, co-author :
Gabelica, Valerie ;  Université de Liège - ULiège
Maeda, R.;  Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Japan
Fujimoto, T.;  Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Japan
Yaku, H.;  Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Japan, Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, 7-1-20 minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan, Advanced Technology Research Laboratories, Panasonic Corporation, 3-4 Hikaridai, Seika-cho, Soraku-gun, Kyoto 619-0237, Japan
Murashima, T.;  Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Japan, Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, 7-1-20 minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan
Sugimoto, N.;  Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Japan, Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, 7-1-20 minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan
Miyoshi, D.;  Faculty of Frontiers of Innovative Research in Science and Technology (FIRST), Japan, Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, 7-1-20 minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan
Title :
Multiple and cooperative binding of fluorescence light-up probe thioflavin t with human telomere DNA G-quadruplex
Publication date :
2013
Journal title :
Biochemistry
ISSN :
0006-2960
eISSN :
1520-4995
Publisher :
American Chemical Society, Washington, United States - District of Columbia
Volume :
52
Issue :
33
Pages :
5620-5628
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 28 October 2016

Statistics


Number of views
117 (4 by ULiège)
Number of downloads
267 (1 by ULiège)

Scopus citations®
 
96
Scopus citations®
without self-citations
90
OpenCitations
 
85

Bibliography


Similar publications



Contact ORBi